Transmembrane serine protease overexpressed in ovarian carcinoma and uses thereof

ABSTRACT

The present invention provides a transmembrane serine protease TADG-12 protein, splice variants of the TADG-12 protein and DNA fragments encoding such proteins. Also provided are vectors and host cells capable of expressing the DNAs. The present invention further provides various methods of early detection and therapies of associated ovarian and other malignancies by utilizing the DNAs and/or proteins disclosed herein.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation-in-part patent application andclaims the benefit of priority under 35 U.S.C. §120 of U.S. Ser. No.09/650,371, filed Aug. 28, 2000, which is a divisional application ofU.S. Ser. No. 09/518,046, now U.S. Pat. No. 6,294,663, which claims thebenefit of priority under 35 U.S.C §120 of U.S. Ser. No. 09/261,416, nowU.S. Pat. No. 6,291,663.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] The present invention relates generally to the fields of cellularbiology and diagnosis of neoplastic disease. More specifically, thepresent invention relates to a transmembrane serine protease termedTumor Associated Differentially-Expressed Gene-12 (TADG-12), which isoverexpressed in ovarian carcinoma.

[0004] 2. Description of the Related Art

[0005] Tumor cells rely on the expression of a concert of proteases tobe released from their primary sites and move to distant sites toinflict lethality. This metastatic nature is the result of an aberrantexpression pattern of proteases by tumor cells and also b y stromalcells surrounding the tumors [1-3]. For most tumors to becomemetastatic, they must degrade their surrounding extracellular matrixcomponents, degrade basement membranes to gain access to the bloodstreamor lymph system, and repeat this process in reverse fashion to settle ina secondary host site [3-6]. All of these processes rely upon what nowappears to be a synchronized protease cascade. In addition, tumor cellsuse the power of proteases to activate growth and angiogenic factorsthat allow the tumor to grow progressively [1]. Therefore, much researchhas been aimed at the identification of tumor-associated proteases andthe inhibition of these enzymes for therapeutic means. More importantly,the secreted nature and/or high level expression of many of theseproteases allows for their detection at aberrant levels in patientserum, e.g. the prostate-specific antigen (PSA), which allows for earlydiagnosis of prostate cancer [7].

[0006] Proteases have been associated directly with tumor growth,shedding of tumor cells and invasion of target organs. Individualclasses of proteases are involved in, but not limited to (1) thedigestion of stroma surrounding the initial tumor area, (2) thedigestion of the cellular adhesion molecules to allow dissociation oftumor cells; and (3) the invasion of the basement membrane formetastatic growth and the activation of both tumor growth factors andangiogenic factors.

[0007] For many forms of cancer, diagnosis and treatment has improveddramatically in the last 10 years. However, the five year survival ratefor ovarian cancer remains below 50% due in large part to the vaguesymptoms which allow for progression of the disease to an advanced stageprior to diagnosis [8]. Although the exploitation of the CA125 antigenhas been useful as a marker for monitoring recurrence of ovarian cancer,it has not proven to be an ideal marker for early diagnosis. Therefore,new markers that may be secreted or released from cells and which arehighly expressed by ovarian tumors could provide a useful tool for theearly diagnosis and for therapeutic intervention in patients withovarian carcinoma.

[0008] The prior art is deficient in the lack of the completeidentification of the proteases overexpressed in carcinoma, therefore,deficient in the lack of a tumor marker useful as an indicator of earlydisease, particularly for ovarian cancers. Specifically, TADG-12, atransmembrane serine protease, has not been previously identified ineither nucleic acid or protein form. The present invention fulfills thislong-standing need and desire in the art.

SUMMARY OF THE INVENTION

[0009] The present invention discloses TADG-12, a new member of theTumor Associated Differentially-Expressed Gene (TADG) family, andvariant splicing forms of TADG-12 (TADG-12V and TADG-12D) that couldlead to a truncated protein product. TADG-12 is a transmembrane serineprotease overexpressed in ovarian carcinoma. The entire cDNA of TADG-12has been identified (SEQ ID No. 1). This sequence encodes a putativeprotein of 454 amino acids (SEQ ID No. 2) which includes a potentialtransmembrane domain, an LDL receptor like domain, a scavenger receptorcysteine rich domain, and a serine protease domain. These features implythat TADG-12 is expressed at the cell surface, and it may be used as amolecular target for therapy or a diagnostic marker.

[0010] The present invention encompasses nucleic acids encoding theTADG-12 protein and its splicing variants, vectors and host cellscapable of expressing the claimed nucleic acids, as well as isolated andpurified TADG-12 and its variant proteins. Specifically, the TADG-12protein and its variants have an amino acid sequence shown in SEQ IDNOs. 2, 4 or 154.

[0011] The present invention further provides methods of diagnosing acancer or malignant hyperplasia in a biological sample b y detecting thepresence of TADG-12 protein or mRNA disclosed herein.

[0012] In another embodiment of the present invention, there is provideda method of targeted therapy by administering a compound having atargeting moiety specific for a TADG-12 protein and a therapeuticmoiety. Specifically, the TADG-12 protein has an amino acid sequenceshown in SEQ ID NOs. 2, 4 or 154.

[0013] Other and further aspects, features, and advantages of thepresent invention will be apparent from the following description of thepresently preferred embodiments of the invention given for the purposeof disclosure.

BRIEF DESCRIPTION OF THE DRAWINGS

[0014] So that the matter in which the above-recited features,advantages and objects of the invention, as well as others which willbecome clear, are attained and can be understood in detail, moreparticular descriptions of the invention briefly summarized above may behad by reference to certain embodiments thereof which are illustrated inthe appended drawings. These drawings form a part of the specification.It is to be noted, however, that the appended drawings illustratepreferred embodiments of the invention and therefore are not to beconsidered limiting in their scope.

[0015]FIG. 1A shows that the expected PCR product of approximately 180bp and the unexpected PCR product of approximately 300 bp using theredundant serine protease primers were not amplified from normal ovarycDNA (Lane 1) but were found in abundance from ovarian tumor cDNA (Lane2). The primer sequences for the PCR reactions are indicated byhorizontal arrows. FIG. 1B shows that TADG-12 was subcloned from the 180bp band while the larger 300 base pair band was designated TADG-12V. Thesequences were found to overlap for 180 base pairs (SEQ ID No. 5 fornucleotide sequence, SEQ ID No. 6 for deduced amino acid sequence) withthe 300 base pair TADG-12V (SEQ ID No. 7 for nucleotide sequence, SEQ IDNo. 8 for deduced amino acid sequence) having an additional insert of133 bases. This insertion (vertical arrow) leads to a frame shift, whichcauses the TADG-12V transcript to potentially produce a truncated formof TADG-12 with a variant amino acid sequence.

[0016]FIG. 2 shows that Northern blot analysis for TADG-12 revealedthree transcripts of 2.4, 1.6 and 0.7 kilobases. These transcripts werefound at significant levels in ovarian tumors and cancer cell lines, butthe transcripts were found only at low levels in normal ovary.

[0017]FIG. 3 shows an RNA dot blot (CLONTECH) probed for TADG-12. Thetranscript was detectable (at background levels) in all 50 of the humantissues represented with the greatest abundance of transcript in theheart. Putamen, amygdala, kidney, liver, small intestine, skeletalmuscle, and adrenal gland were also found to have intermediate levels ofTADG-12 transcript.

[0018]FIG. 4 shows the entire cDNA sequence for TADG-12 (SEQ ID No. 1)with its predicted open reading frame of 454 amino acids (SEQ ID No. 2).Within the nucleotide sequence, the Kozak's consensus sequence for theinitiation of translation and the poly-adenylation signal areunderlined. In the protein sequence, a potential transmembrane domain isboxed. The low-density lipoprotein receptor-like class A domain isunderlined with a solid line. The scavenger receptor cysteine richdomain (SRCR) domain is underlined with a broken line. The residues ofthe catalytic triad of the serine protease domain are circled, and thebeginning of the catalytic domain is marked with an arrow designated asa potential proteolytic cleavage site. The asterik represents the stopcodon that terminates translation.

[0019]FIG. 5A shows the 35 amino acid low-density lipoproteinreceptor-like class A domain of TADG-12 (SEQ ID No. 13) aligned withother low-density lipoprotein receptor-like class A domain motifs fromthe serine protease TMPRSS2 (U75329, SEQ ID No. 14), the complementsubunit C8 (P07358, SEQ ID No. 9), two LDLR-A domains of theglycoprotein GP300 (P98164, SEQ ID Nos. 11-12), and the serine proteasematriptase (AF118224, SEQ ID No. 10). TADG-12 has its highest similaritywith the other serine proteases for which it is 54% similar to TMPRSS2and 53% similar to matriptase. The highly conserved cysteine residuesare shown in bold type.

[0020]FIG. 5B shows the scavenger receptor cysteine rich domain ofTADG-12 (SEQ ID No. 17) aligned with other domain family membersincluding the human macrophage scavenger receptor (P21757, SEQ ID No.16), human enterokinase (P98073, SEQ ID No. 19), bovine enterokinase(P21758, SEQ ID No. 15), and the serine protease TMPRSS2 (SEQ ID No.18). Again, TADG-12 shows its highest similarity within this region tothe protease TMPRSS2 at 43%.

[0021]FIG. 5C shows the protease domain of TADG-12 (SEQ ID No. 23) inalignment with other human serine proteases including protease M(U62801, SEQ ID No. 20), trypsinogen I (P07477, SEQ ID No. 21), plasmakallikrein (P₀₃₉₅₂, SEQ ID No. 22), hepsin (P₀₅₉₈₁, SEQ ID No. 25), andTMPRSS2 (SEQ ID No. 24). Cons represents the consensus sequence for eachalignment.

[0022]FIG. 6 shows semi-quantitative PCR analysis that was performed forTADG-12 (upper panel) and TADG-12V (lower panel). The amplification ofTADG-12 or TADG-12V was performed in parallel with PCR amplification ofβ-tubulin product as an internal control. The TADG-12 transcript wasfound to be overexpressed in 41 of 55 carcinomas. The TADG-12Vtranscript was found to be overexpressed in 8 of 22 carcinomas examined.Note that the samples in the upper panel are not necessarily the same asthe samples in the lower panel.

[0023]FIG. 7 shows immunohistochemical staining of normal ovary andovarian tumors which were performed using a polyclonal rabbit antibodydeveloped to a TADG-12 specific peptide. No significant staining wasdetected in normal ovary (FIG. 7A). Strong positive staining wasobserved in 22 of 29 carcinomas examined. FIGS. 7B and 7C represent aserous and mucinous carcinoma, respectively. Both show diffuse stainingthroughout the cytoplasm of tumor cells while stromal cells remainrelatively unstained.

[0024]FIG. 8 is a model to demonstrate the progression of TADG-12 withina cellular context. In normal circumstances, the TADG-12 transcript isappropriately spliced and the resulting protein is capable of beingexpressed at the cell surface where the protease may be cleaved to anactive form. The role of the remaining ligand binding domains has notyet been determined, but one can envision their potential to bind othermolecules for activation, internalization or both. The TADG-12Vtranscript, which occurs in some tumors, may be the result of mutationand/or poor mRNA processing may b e capable of producing a truncatedform of TADG-12 that does not have a functional protease domain. Inaddition, this truncated product may present a novel epitope at thesurface of tumor cells.

[0025]FIG. 9 shows the schematic for TADG-12 and splicing variantsTADG-12D and TADG-12V. TM, transmembrane domain; LDLR, low-densitylipoprotein receptor-like domain; SRCR, scavenger receptor cysteine richdomain.

[0026]FIG. 10 compares the protease domain of TADG-12 and splicingvariants TADG-12D and TADG-12V.

[0027]FIG. 11 shows the expression of TADG-12D in normal and carcinomatissue.

[0028]FIG. 12 shows the expression of TADG-12D in various carcinomatissue.

DETAILED DESCRIPTION OF THE INVENTION

[0029] The present invention discloses a serine protease identifiedusing a PCR based strategy. By Northern blot, the largest transcript forthis gene is approximately 2.4 kb, and it was found to be expressed athigh levels in ovarian tumors while found at minimal levels in all othertissues examined. The full-length cDNA encoding a novel multi-domain,cell-surface serine protease named TADG-12 was cloned. The TADG-12 cDNAis 2413 base pairs long (SEQ ID NO. 1) encoding a 454 amino acid protein(SEQ ID NO. 2). A variant form, TADG-12V (SEQ ID NO. 3), encodes a 294amino acid protein (SEQ ID NO. 4). Another variant form, a 344 aminoacid protein TADG-12D (SEQ ID NO. 154) was encoded by the DNA of SEQ IDNO. 155.

[0030] The TADG-12 protein contains a cytoplasmic domain, a type IItransmembrane domain, a low-density lipoprotein receptor-like class Adomain, an scavenger receptor cysteine rich domain and a serine proteasedomain. Using a semi-quantitative PCR analysis, it was shown thatTADG-12 was overexpressed in a majority of tumors studied.Immunohistochemical staining corroborates that in some cases thisprotein is localized to the cell-surface of tumor cells and thissuggests that TADG-12 has some extracellular proteolytic functions. Thevariant splicing form TADG-12V is present in 35% of the tumors studied.Another splicing variant TADG-12D is also highly expressed in tumorcells. These variant mRNAs would lead to truncated proteins that mayprovide unique peptide sequences on the surface of tumor cells.

[0031] The TADG-12 protein contains two extracellular domains whichmight confer unusual properties to this multidomain molecule. Althoughthe precise role of low-density lipoprotein receptor-like class A domainfunction with regard to proteases remains unclear, this domain certainlyhas the capacity to bind calcium and other positively charged ligands[9, 10]. This may play an important role in the regulation of theprotease or subsequent internalization of the molecule. The scavengerreceptor cysteine rich domain was originally identified within themacrophage scavenger receptor and functionally described to bindlipoproteins. Not only are scavenger receptor cysteine rich domainscapable of binding lipoproteins, but they may also bind to molecules asdiverse as polynucleotides [11]. More recent studies have identifiedmembers of this domain family in proteins with functions that vary fromproteases to cell adhesion molecules involved in maturation of theimmune system [12]. In addition, TADG-12, like TMPRSS2 has only four ofsix cysteine residues conserved within its scavenger receptor cysteinerich domain. This difference may allow for different structural featuresof these domains that confer unusual ligand binding properties.

[0032] At this time, only the function of the CD6 encoded scavengerreceptor cysteine rich domain is well documented. In the case of CD6,the scavenger receptor cysteine rich domain binds to the cell adhesionmolecule ALCAM [11]. This mediation of cell adhesion is a usefulstarting point for future research on newly identified scavengerreceptor cysteine rich domains; however, the possibility of multiplefunctions for this domain can not be overlooked. Scavenger receptorcysteine rich domains are certainly capable of cell adhesion typeinteractions, but their capacity to bind other types of ligands shouldbe considered.

[0033]FIG. 8 presents a working model of TADG-12 with the informationdisclosed in the present invention. Two transcripts are produced whichlead to the production of either TADG-12 or the truncated TADG-12Vproteins. Either of these proteins is potentially targeted to the cellsurface. TADG-12 is capable of becoming an activated serine proteasewhile TADG-12V is a truncated protein product that if at the cellsurface may represent a tumor specific epitope.

[0034] The problem with treatment of cancer such as ovarian cancerremains the inability to diagnose the disease at an early stage.Identifying genes that are expressed early in the disease process suchas proteases that are essential for tumor cell growth [13] is animportant step toward improving treatment. The availability of theTADG-12 and its splicing variants opens the way for a number studiesthat can lead to various applications. For example, the TADG-12 and itsvariant genes or proteins can be used as a diagnostic or therapeutictarget in ovarian and other carcinomas. The extracellular ligand bindingdomains are natural targets for drug delivery systems, and aberrantpeptide associated with the TADG-12 protein or its variants may provideexcellent targets for immune stimulation. Alternatively, inhibition ofenzymes such as TADG-12 may be an effective means for slowingprogression of various cancer and improving the quality of patient life.

[0035] As described herein, the invention provides a number ofdiagnostic advantages and uses. For example, the TADG-12 protein or itsvariants disclosed in the present invention are useful in diagnosingcancer in different tissues since these proteins are highly expressed intumor cells. Antibodies (or antigen-binding fragments thereof) whichbind to an epitope specific for a TADG-12 protein or its variantproteins are useful in a method of detecting TADG-12 or its variants ina biological sample for diagnosis of cancerous or neoplastictransformation. This method includes the steps of obtaining a biologicalsample (e.g., cells, blood, plasma, tissue, etc.) from a patientsuspected of having cancer, contacting the sample with a labeledantibody (e.g., radioactively tagged antibody) specific for TADG-12 orits splicing variants, and detecting the proteins using standardimmunoassay techniques. Likewise, a standard Northern blot assay can beused to ascertain the relative amounts of TADG-12 mRNA in a cell ortissue obtained from a patient suspected of having cancer in accordancewith conventional Northern hybridization techniques known to those ofordinary skill in the art.

[0036] In accordance with the present invention there may b e employedconventional molecular biology, microbiology, and recombinant DNAtechniques within the skill of the art. Such techniques are explainedfully in the literature. See, e.g., Maniatis, Fritsch & Sambrook,“Molecular Cloning: A Laboratory Manual (1982); “DNA Cloning: APractical Approach,” Volumes I and II (D. N. Glover ed. 1985);“Oligonucleotide Synthesis” (M. J. Gait ed. 1984); “Nucleic AcidHybridization” [B. D. Hames & S. J. Higgins eds. (1985)]; “Transcriptionand Translation” [B. D. Hames & S. J. Higgins eds. (1984)]; “Animal CellCulture” [R. I. Freshney, ed. (1986)]; “Immobilized Cells And Enzymes”[IRL Press, (1986)]; B. Perbal, “A Practical Guide To Molecular Cloning”(1984).

[0037] As used herein, “TADG-12 protein” refers to full length TADG-12protein (SEQ ID NO. 2) as well as splicing variants or truncated formsof TADG-12. These splicing variants or truncated forms are derived fromTADG-12 by removing or deleting amino acids from SEQ ID NO. 2 with orwithout inclusion of addition amino acids. Examples of TADG-12 splicingvariants include, but are not limited to, TADG-12V and TADG-12D.

[0038] The present invention is directed to DNA fragments encodingTADG-12 and its splicing variants (such as TADG-12V and TADG-12D). Theclaimed DNAs include those having different codon sequences for TADG-12due to the degeneracy of the genetic code. The protein encoded by theDNA of this invention may share at least 80% sequence identity(preferably 85%, more preferably 90%, and most preferably 95%) with theamino acids listed in SEQ ID No. 2 or SEQ ID No. 4. More preferably, theDNA includes the coding sequence of SEQ ID NO. 1, or a degeneratevariant of such a sequence. Preferably, the DNA has the sequence shownin SEQ ID NOs. 1, 3 or 155, and the DNA encodes a TADG-12 protein havingthe amino acid sequence shown in SEQ ID NOs. 2, 4 or 154.

[0039] This invention includes a substantially pure DNA comprising asequence of at least 15 consecutive nucleotides (preferably 20, morepreferably 30, even more preferably 50, and most preferably all) of theregion from nucleotides 1 to 2413 of the nucleotides listed in SEQ IDNo. 1, or of the region from nucleotides 1 to 2544 of the nucleotideslisted in SEQ ID No. 3. The present invention also comprises antisenseoligonucleotides directed against DNAs encoding TADG-12 (SEQ ID NO. 1)or its splicing variants (e.g. SEQ ID Nos. 3 or 155). Given theteachings of the present invention, a person having ordinary skill inthis art would readily be able to develop antisense oligonucleotidesdirected against these DNA sequences.

[0040] The present invention also provides a vector comprising a DNAsequence which encodes a human TADG-12 protein or its splicing variants.The vector comprising in operable linkage an origin of replication, apromoter and a DNA sequence coding for said protein is capable ofreplication in a host cell. Preferably, the vector contains a portion ofthe DNA sequences shown in SEQ ID NOs. 1, 3 or 155. Methods which arewell known to those skilled in the art can be used to constructexpression vectors containing appropriate transcriptional andtranslational control signals. See for example, the techniques describedin Sambrook et al., 1989, Molecular Cloning: A Laboratory Manual (2ndEd.), Cold Spring Harbor Press, N.Y. Vectors of the invention include,but are not limited to, plasmid vectors and viral vectors. Preferredviral vectors of the invention are those derived from retroviruses,adenovirus, adeno-associated virus, SV40 virus, or herpes viruses.Representative host cells include bacterial cells, yeast cells,mammalian cells and insect cells.

[0041] The present invention is also directed to an isolated andpurified TADG-12 protein encoded for by the DNA claimed herein.Preferably, the isolated and purified TADG-12 protein has the amino acidsequence shown in SEQ ID NOs 2, 4 or 154.

[0042] By a “substantially pure protein” is meant a protein which hasbeen separated from at least some of those components which naturallyaccompany it. A substantially pure TADG-12 protein may be obtained, forexample, by extraction from a natural source, b y expression of arecombinant nucleic acid encoding a TADG-12 polypeptide, or bychemically synthesizing the protein.

[0043] Included in this invention are TADG-12 proteins which are encodedat least in part by portions of SEQ ID No. 1 or SEQ ID No. 3, e.g.,products of alternative mRNA splicing or alternative protein processingevents, or in which a section of TADG-12 sequence has been deleted. Thefragment, or the intact TADG-12 polypeptide, may be covalently linked toanother polypeptide, e.g. which acts as a label, a ligand or a means toincrease antigenicity.

[0044] In addition to substantially full-length proteins, the inventionalso includes fragments (e.g., antigenic fragments) of the TADG-12protein. As used herein, “fragment,” as applied to a polypeptide, willordinarily be at least 10 residues, more typically at least 20 residues,and preferably at least 30 (e.g., 50) residues in length, but less thanthe entire, intact sequence. Fragments of the TADG-12 protein can begenerated by methods known to those skilled in the art, e.g., byenzymatic digestion of naturally occurring or recombinant TADG-12protein, by recombinant DNA techniques using an expression vector thatencodes a defined fragment of TADG-12, or by chemical synthesis. Theability of a candidate fragment to exhibit a characteristic of TADG-12(e.g., binding to an antibody specific for TADG-12) can be assessed bymethods described herein. Purified TADG-12 or antigenic fragments ofTADG-12 can be used to generate new antibodies or to test existingantibodies (e.g., as positive controls in a diagnostic assay) byemploying standard protocols known to those skilled in the art.

[0045] The invention also includes a polyclonal or monoclonal antibodywhich specifically binds to TADG-12 protein or its variants. Standardprotocols for monoclonal and polyclonal antibody production known tothose skilled in this art are employed. The invention encompasses notonly intact monoclonal antibody, but also immunologically-activeantibody fragment, e.g., a Fab or (Fab)₂ fragment, an engineered singlechain Fv molecule, or a chimeric antibody consists of regions derivedfrom different species (e.g., the antigen binding sites are of murineorigin and the remaining portions of the antibody are of human origin).

[0046] The anti-TADG-12 antibody, or a fragment thereof, may be linkedto a toxin or to a detectable label, e.g. a radioactive label,non-radioactive isotopic label, fluorescent label, chemiluminescentlabel, paramagnetic label, enzyme label, or calorimetric label. One ofordinary skill in the art would know of suitable labels which may beemployed in accordance with the present invention. The binding of theselabels to antibodies or fragments thereof can be accomplished usingstandard techniques commonly known to those of ordinary skill in theart. Typical techniques are described by Kennedy et al., (1976) Clin.Chim. Acta 70:1-31; and Schurs et al., (1977) Clin. Chim. Acta 81:1-40.

[0047] In another embodiment of the present invention, there areprovided a methods for detecting malignant hyperplasia by detecting aTADG-12 protein or its variants, or mRNA encoding these proteins in abiological sample. Preferably, the biological sample is selected fromthe group consisting of blood, urine, saliva, tears, interstitial fluid,ascites fluid, tumor tissue biopsy and circulating tumor cells. TADG-12protein or mRNA can be detected by Northern blot, Western blot, PCR, dotblot, ELISA sandwich assay, radioimmunoassay, DNA array chips or flowcytometry. Such methods can be used for detecting ovarian cancer, breastcancer, lung cancer, colon cancer, prostate cancer and other cancers inwhich TADG-12 is overexpressed.

[0048] In still another embodiment of the present invention, there isprovided a method of inhibiting expression of endogenous TADG-12 mRNA ina cell by introducing a vector comprising a DNA fragment of TADG-12 inopposite orientation operably linked to elements necessary forexpression. As a result, the vector produces TADG-12 antisense mRNA inthe cell, which hybridizes to endogenous TADG-12 mRNA, therebyinhibiting expression of endogenous TADG-12 mRNA.

[0049] In still yet another embodiment of the present invention, thereis provided a method of inhibiting expression of a TADG-12 protein byintroducing an antibody directed against a TADG-12 protein or fragmentthereof. As a result, the binding of the antibody to the TADG-12 proteinor fragment thereof inhibits the expression of the TADG-12 protein.

[0050] In another embodiment of the present invention, TADG-12 geneproducts including the truncated form can be used for targeted therapy.Specifically, a compound having a targeting moiety specific for aTADG-12 protein or its variants and a therapeutic moiety is administeredto an individual in need of such treatment. Preferably, the targetingmoiety is selected from the group consisting of a n antibody directedagainst a TADG-12 protein or its variants and a ligand or ligand bindingdomain that binds a TADG-12 protein. For example, the TADG-12 proteincan have an amino acid sequence shown in SEQ ID NOs. 2, 4 or 154. Stillpreferably, the therapeutic moiety is selected from the group consistingof a radioisotope, a toxin, a chemotherapeutic agent, an immunestimulant and a cytotoxic agent. Such a method can be used for treatingan individual having, for example, ovarian cancer, lung cancer, prostatecancer, or colon cancer.

[0051] In yet another embodiment of the present invention, there isprovided a method of vaccinating, or producing an immune response in, anindividual against TADG-12 by inoculating the individual with a TADG-12protein or fragment thereof. Specifically, the TADG-12 protein orfragment thereof lacks TADG-12 activity, and the inoculation elicits animmune response in the individual, thereby vaccinating the individualagainst TADG-12. Preferably, the individual has a cancer, is suspectedof having a cancer or is at risk of getting a cancer. Still preferably,TADG-12 protein has an amino acid sequence shown in SEQ ID NOs. 2, 4 or154, while TADG-12 fragment has a sequence shown in SEQ ID No. 8, or isa 9-residue fragment up to a 20-residue fragment. Examples of 9-residuefragment are shown in SEQ ID Nos. 35, 36, 55, 56, 83, 84, 97, 98, 119,120, 122, 123 and 136.

[0052] In still yet another embodiment of the present invention, thereis provided an immunogenic composition, comprising a n immunogenicfragment of a TADG-12 protein and an appropriate adjuvant. Preferably,the immunogenic fragment of the TADG-12 protein has a sequence shown inSEQ ID No. 8, or is a 9-residue fragment up to a 20-residue fragment.Examples of 9-residue fragment are shown in SEQ ID Nos. 35, 36, 55, 56,83, 84, 97, 98, 119, 120, 122, 123 and 136.

[0053] The following examples are given for the purpose of illustratingvarious embodiments of the invention and are not meant to limit thepresent invention in any fashion.

EXAMPLE 1

[0054] Tissue Collection and Storage

[0055] Upon patient hysterectomy, bilateral salpingo-oophorectomy, orsurgical removal of neoplastic tissue, the specimen is retrieved andplaced on ice. The specimen was then taken to the resident pathologistfor isolation and identification of specific tissue samples. Finally,the sample was frozen in liquid nitrogen, logged into the laboratoryrecord and stored at −80° C. Additional specimens were frequentlyobtained from the Cooperative Human Tissue Network (CHTN). These sampleswere prepared by the Cooperative Human Tissue Network and shipped on dryice. Upon arrival, these specimens were logged into the laboratoryrecord and stored at −80° C.

EXAMPLE 2

[0056] mRNA Extraction and cDNA Synthesis

[0057] Sixty-nine ovarian tumors (4 benign tumors, 10 low malignantpotential tumors and 55 carcinomas) and 10 normal ovaries were obtainedfrom surgical specimens and frozen in liquid nitrogen. The human ovariancarcinoma cell lines SW 626 and Caov 3, the human breast carcinoma celllines MDA-MB-231 and MDA-MB-435S were purchased from the American TypeCulture Collection (Rockville, Md.). Cells were cultured tosub-confluency in Dulbecco's modified Eagle's medium, supplemented with10% (v/v) fetal bovine serum and antibiotics.

[0058] Extraction of mRNA and cDNA synthesis were carried out by themethods described previously [14-16]. mRNA was isolated by using aRiboSep mRNA isolation kit (Becton Dickinson Labware). In thisprocedure, poly A+ mRNA was isolated directly from the tissue lysateusing the affinity chromatography media oligo(dT) cellulose. cDNA wassynthesized with 5.0 μg of mRNA by random hexamer priming using 1ststrand cDNA synthesis kit (CLONTECH).

EXAMPLE 3

[0059] PCR with Redundant Primers and Cloning of TADG-12 cDNA

[0060] Redundant primers, forward 5′-TGGGTIGTIACIGCIG CICA(CT)TG-3′ (SEQID No. 26) and reverse 5′-A(AG)IA(AG)IGCIATITCI TTICC-3′ (SEQ ID No.27), for the consensus sequences of amino acids surrounding thecatalytic triad for serine proteases were used to compare the PCRproducts from normal and carcinoma cDNAs. The appropriate bands wereligated into Promega T-vector plasmid and the ligation product was usedto transform JM109 cells (Promega) grown on selection media. Afterselection of individual colonies, they were cultured and plasmid DNA wasisolated by means of the Wizard miniprep DNA purification system(Promega). Nucleotide sequencing was performed using PRISM ReadyReaction Dye Deoxy terminator cycle sequencing kit (Applied Biosystems).Applied Biosystems Model 373A DNA sequencing system was used for directcDNA sequence determination.

[0061] The original TADG-12 subclone was randomly labeled and used as aprobe to screen an ovarian tumor cDNA library by standard hybridizationtechniques [15, 17]. The library was constructed in λZAP using mRNAisolated from the tumor cells of a stage III/grade III ovarianadenocarcinoma patient. Three overlapping clones were obtained whichspanned 2315 nucleotides. The final 99 nucleotides encoding the most 3′sequence including the poly A tail was identified by homology withclones available in the GenBank EST database.

EXAMPLE 4

[0062] Quantitative PCR

[0063] The mRNA overexpression of TADG-12 was determined using aquantitative PCR. Quantitative PCR was performed according to theprocedure as previously reported [16]. Oligonucleotide primers were usedfor: TADG-12, forward 5′-GAAACATGTCCTTGCTCTCG-3′ (SEQ ID No. 28) andreverse 5′-ACTAACTTCCACAGCCTCCT-3′ (SEQ ID No. 29); the variant TADG-12,forward 5′-TCCAGGTGGGTCAGTTTCC-3′ (SEQ ID No. 30), reverse5′-CTCTTGGCTTGTACTTGCT-3′ (SEQ ID No. 31); β-tubulin, forward5′-CGCATCAACGTGTACTACAA-3′ (SEQ ID No. 32) and reverse5′-TACGAGCTGGTGGACTGAGA-3′ (SEQ ID No. 33). β-tubulin was utilized as aninternal control. The PCR reaction mixture consists of cDNA derived from50 ng of mRNA, 5 pmol of sense and antisense primers for both theTADG-12 gene and the β-tubulin gene, 200 μmol of dNTPs, 5 μCi of α-³²PdCTP and 0.25 unit of Taq DNA polymerase with reaction buffer (Promega)in a final volume of 25 μl. The target sequences were amplified inparallel with the β-tubulin gene. Thirty cycles of PCR were carried outin a Thermal Cycler (Perkin-Elmer Cetus). Each cycle of PCR included 30seconds of denaturation at 94%C, 30 seconds of annealing at 60%C and 30seconds of extension at 72%C. The PCR products were separated on 2%agarose gels and the radioactivity of each PCR product was determined byusing a Phospho Imager (Molecular Dynamics). The present study used theexpression ratio (TADG-12/β-tubulin) as measured b y phosphoimager toevaluate gene expression and defined the value at mean+2SD of normalovary as the cut-off value to determine overexpression. The student's ttest was used for comparison of the mean values of normal ovary andtumors.

EXAMPLE 5

[0064] Sequencing of TADG-12 Utilizing a plasmid specific primer nearthe cloning site, sequencing reactions were carried out using PRISM™Ready Reaction Dye Deoxy™ terminators (Applied Biosystems cat# 401384)according to the manufacturer's instructions. Residual dye terminatorswere removed from the completed sequencing reaction using a Centri-sep™spin column (Princeton Separation cat.# CS-901). An Applied BiosystemsModel 373A DNA Sequencing System was available and was used for sequenceanalysis.

EXAMPLE 6

[0065] Antibody Production

[0066] Polyclonal rabbit antibodies were generated b y immunization ofwhite New Zealand rabbits with a poly-lysine linked multiple antigenpeptide derived from the TADG-12 carboxy-terminal protein sequenceNH₂-WIHEQMERDLKT-COOH (WIHEQMERDLKT, SEQ ID No. 34). This peptide ispresent in full length TADG-12, but not TADG-12V. Rabbits were immunizedwith approximately 100 μg of peptide emulsified in Ribi adjuvant.Subsequent boost immunizations were carried out at 3 and 6 weeks, andrabbit serum was isolated 10 days after the boost inoculations. Serawere tested by dot blot analysis to determine affinity for the TADG-12specific peptide. Rabbit pre-immune serum was used as a negativecontrol.

EXAMPLE 7

[0067] Northern Blot Analysis

[0068] 10 μg of mRNA were loaded onto a 1% formaldehyde-agarose gel,electrophoresed and blotted on a Hybond-N+ nylon membrane (Amersham).³²P-labeled cDNA probes were made by Prime-a-Gene Labeling System(Promega). The PCR products amplified by the same primers as above wereused for probes. The blots were prehybridized for 30 min and hybridizedfor 60 min at 68%C with ³²P-labeled cDNA probe in ExpressHybHybridization Solution (CLONTECH). Control hybridization to determinerelative gel loading was performed with the β-tubulin probe.

[0069] Normal human tissues; spleen, thymus, prostate, testis, ovary,small intestine, colon and peripheral blood leukocyte, and normal humanfetal tissues; brain, lung, liver and kidney (Human Multiple TissueNorthern Blot; CLONTECH) were also examined b y same hybridizationprocedure.

EXAMPLE 8

[0070] Immunohistochemistry

[0071] Immunohistochemical staining was performed using a VectastainElite ABC Kit (Vector). Formalin fixed and paraffin embedded specimenswere routinely deparaffinized and processed using microwave heattreatment in 0.01 M sodium citrate buffer (pH 6.0). The specimens wereincubated with normal goat serum in a moist chamber for 30 minutes.TADG-12 peptide antibody was allowed to incubate with the specimens in amoisture chamber for 1 hour. Excess antibody was washed away withphosphate buffered saline. After incubation with biotinylatedanti-rabbit IgG for 30 minutes, the sections were then incubated withABC reagent (Vector) for 30 minutes. The final products were visualizedusing the AEC substrate system (DAKO) and sections were counterstainedwith hematoxylin before mounting. Negative controls were performed b yusing normal serum instead of the primary antibody.

EXAMPLE 9

[0072] Isolation of Catalytic Domain Subclones of TADG-12 and TADG-12Variant

[0073] To identify serine proteases that are expressed in ovariantumors, redundant PCR primers designed to the conserved regions of thecatalytic triad of these enzymes were employed. A sense primer designedto the region surrounding the conserved histidine and an anti-senseprimer designed to the region surrounding the conserved aspartate wereused in PCR reactions with either normal ovary or ovarian tumor cDNA astemplate. In the reaction with ovarian tumor cDNA, a strong product bandof the expected size of approximately 180 bp was observed as well as anunexpected PCR product of approximately 300 bp which showed strongexpression in some ovarian tumor cDNA's (FIG. 1A). Both of these PCRproducts were subcloned and sequenced. The sequence of the subclonesfrom the 180 bp band (SEQ ID No. 5) was found to be homologous to thesequence identified in the larger, unexpected band (SEQ ID No. 7) exceptthat the larger band had an additional insert of 133 nucleotides (FIG.1B). The smaller product of the appropriate size encoded for a proteinsequence (SEQ ID No. 6) homologous to other known proteases while thesequence with the insertion (SEQ ID No. 8) encoded for a frame shiftfrom the serine protease catalytic domain and a subsequent prematuretranslational stop codon. TADG-12 variants from four individual tumorswere also subcloned and sequenced. It was found that the sequence andinsert to be identical. The genomic sequences for these cDNA derivedclones were amplified by PCR, examined and found to contain potentialAG/GT splice sites that would allow for the variant transcriptproduction (data not shown).

EXAMPLE 10

[0074] Northern Blot Analysis of TADG-12 Expression

[0075] To examine transcript size and tissue distribution, the catalyticdomain subclone was randomly labeled and used to probe Northern blotsrepresenting normal ovarian tissue, ovarian tumors and the cancer celllines SW626, CAOV3, HeLa, MD-MBA-435S and MD-MBA-231 (FIG. 2). Threetranscripts of 2.4, 1.6 and 0.7 kilobases were observed. In blots ofnormal and ovary tumor the smallest transcript size 0.7 kb was lowlyexpressed in normal ovary while all transcripts (2.4, 1.6 and 0.7 kb)were abundantly present in serous carcinoma. In addition, Northern blotsrepresenting the normal human tissues spleen, thymus, prostate, testis,ovary, small intestine, colon and peripheral blood leukocyte, and normalhuman fetal tissues of brain, lung, liver and kidney were examined. Thesame three transcripts were found to be expressed weakly in all of thesetissues (data not shown). A human β-tubulin specific probe was utilizedas a control for relative sample loading. In addition, an RNA dot blotwas probed representing 50 human tissues and determined that this cloneis weakly expressed in all tissues represented (FIG. 3). It was foundmost prominently in heart, with intermediate levels in putamen,amygdala, kidney, liver, small intestine, skeletal muscle, and adrenalgland.

EXAMPLE 11

[0076] Sequencing and Characterization of TADG-12

[0077] An ovarian tumor cDNA library constructed in λZAP was screened bystandard hybridization techniques using the catalytic domain subclone asa probe. Two clones that overlapped with the probe were identified andsequenced and found to represent 2316 nucleotides. The 97 nucleotides atthe 3′ end of the transcript including the poly-adenylation signal andthe poly (A) tail were identified by homology with clones available inGenBank's EST database. This brought the total size of the transcript to2413 bases (SEQ ID No. 1, FIG. 4). Subsequent screening of GenBank'sGenomic Database revealed that TADG-12 is homologous to a cosmid fromchromosome 17. This cosmid has the accession number AC015555.

[0078] The identified cDNA includes an open reading frame that wouldproduce a predicted protein of 454 amino acids (SEQ ID No. 2), namedTumor Associated Differentially-Expressed Gene 12 (TADG-12). Thesequence has been submitted to the GenBank database and granted theaccession #AF201380. Using homology alignment programs, this proteincontains several domains including an amino-terminal cytoplasmic domain,a potential Type II transmembrane domain followed by a low-densitylipoprotein receptor-like class A domain (LDLR-A), a scavenger receptorcysteine rich domain (SRCR), and an extracellular serine proteasedomain.

[0079] As predicted by the ™Pred program, TADG-12 contains a highlyhydrophobic stretch of amino acids that could serve as a potentialtransmembrane domain, which would retain the amino terminus of theprotein within the cytoplasm and expose the ligand binding domains andprotease domain to the extracellular space. This general structure isconsistent with other known transmembrane proteases including hepsin[18], and TMPRSS2 [19], and TADG-12 is particularly similar in structureto the TMPRSS2 protease.

[0080] The low-density lipoprotein receptor-like class A domain ofTADG-12 is represented by the sequence from amino acid 74 to 108 (SEQ IDNo. 13). The low-density lipoprotein receptor-like class A domain wasoriginally identified within the LDL Receptor [20] as a series ofrepeated sequences of approximately 40 amino acids, which contained 6invariant cysteine residues and highly conserved aspartate and glutamateresidues. Since that initial identification, a host of other genes havebeen identified which contain motifs homologous to this domain [21].Several proteases have been identified which contain LDLR-A motifsincluding matriptase, TMPRSS2 and several complement components. Acomparison of TADG-12 with other known low-density lipoproteinreceptor-like class A domains is shown in FIG. 5A. The similarity ofthese sequences range from 44 to 54% of similar or identical aminoacids.

[0081] In addition to the low-density lipoprotein receptor-like class Adomain, TADG-12 contains another extracellular ligand binding domainwith homology to the group A scavenger receptor cysteine rich domainfamily. This family of protein domains typically is defined by theconservation of 6 cysteine resides within a sequence of approximately100 amino acids [11]. The scavenger receptor cysteine rich domain ofTADG-12 is encoded by amino acids 109 to 206 (SEQ ID No. 17), and thisdomain was aligned with other scavenger receptor cysteine rich domainsand found to have between 36 and 43% similarity (FIG. 5B). However,TADG-12 only has 4 of the 6 conserved cysteine residues. This is similarto the scavenger receptor cysteine rich domain found in the proteaseTMPRSS2.

[0082] The TADG-12 protein also includes a serine protease domain of thetrypsin family of proteases. An alignment of the catalytic domain ofTADG-12 with other known proteases is shown in FIG. 5C. The similarityamong these sequence ranges from 48 to 55%, and TADG-12 is most similarto the serine protease TMPRSS2 which also contains a transmembranedomain, low-density lipoprotein receptor-like class A domain and anscavenger receptor cysteine rich domain. There is a conserved amino acidmotif (RIVGG) downstream from the scavenger receptor cysteine richdomain that is a potential cleavage/activation site common to manyserine proteases of this family [22]. This suggests that TADG-12 istrafficked to the cell surface where the ligand binding domains arecapable of interacting with extracellular molecules and the proteasedomain is potentially activated. TADG-12 also contains conservedcysteine residues (amino acids 208 and 243) which in other proteasesform a disulfide bond capable of linking the activated protease to theother extracellular domains.

EXAMPLE 12

[0083] Quantitative PCR Characterization of TADG-12V

[0084] The original TADG-12 subclone was identified as highly expressedin the initial redundant-primer PCR experiment. The TADG-12 variant form(TADG-12V) with the insertion of 133 bp was also easily detected in theinitial experiment. To identify the frequency of this expression andwhether or not the expression level between normal ovary and ovariantumors was different, a previously authenticated semi-quantitative PCRtechnique was employed [16]. The PCR analysis co-amplified a product forβ-tubulin with either a product specific to TADG-12 or TADG-12V in thepresence of a radiolabelled nucleotide. The products were separated byagarose gel electrophoresis and a phosphoimager was used to quantitatethe relative abundance of each PCR product.

[0085] Examples of these PCR amplification products are shown for bothTADG-12 and TADG-12V in FIG. 6. Normal expression was defined as themean ratio of TADG-12 (or TADG-12V) to β-tubulin +/−2SD as examined innormal ovarian samples. For tumor samples, overexpression was definedas >2SD from the normal TADG-12/β-tubulin or TADG-12V/β-tubulin ratio.The results are summarized in Table 1 and Table 2. TADG-12 was found tobe overexpressed in 41 of 55 carcinomas examined while the variant formwas present a t aberrantly high levels in 8 of 22 carcinomas. Asdetermined by the student's t test, these differences were statisticallysignificant (p<0.05). TABLE 1 Frequency of Overexpression of TADG-12 inOvarian Carcinoma Histology Type TADG-12 (%) Normal  0/16 (0%)LMP-Serous  3/6 (50%) LMP-Mucinous  0/4 (0%) Serous Carcinoma 23/29(79%) Mucinous Carcinoma  7/12 (58%) Endometrioid Carcinoma  8/8 (100%)Clear Cell Carcinoma  3/6 (50%) Benign Tumors  3/4 (75%)

[0086] Overexpression: more than two standard deviations above the meanfor normal ovary.

[0087] LMP low malignant potential tumor. TABLE 2 Frequency ofOverexpression of TADG-12V in Ovarian Carcinoma Histology Type TADG-12V(%) Normal 0/10 (0%) LMP-Serous 0/5 (0%) LMP-Mucinous 0/3 (0%) SerousCarcinoma 4/14 (29%) Mucinous Carcinoma 3/5 (60%) Endometrioid Carcinoma1/3 (33%) Clear Cell Carcinoma N/D

[0088] Overexpression: more than two standard deviations above the meanfor normal ovary; LMP low malignant potential tumor

EXAMPLE 13

[0089] Immunohistochemical Analysis of TADG-12 in Ovarian Tumor Cells

[0090] In order to examine the TADG-12 protein, polyclonal rabbitanti-sera to a peptide located in the carboxy-terminal amino acidsequence was developed. These antibodies were used to examine theexpression level of the TADG-12 protein and its localization withinnormal ovary and ovarian tumor cells by immuno-localization. No stainingwas observed in normal ovarian tissues (FIG. 7A) while significantstaining was observed in 22 of 29 tumors studied. Representative tumorsamples are shown in FIGS. 7B and 7C. It should be noted that TADG-12 isfound in a diffuse pattern throughout the cytoplasm indicative of aprotein in a trafficking pathway. TADG-12 is also found at the cellsurface in these tumor samples as expected. It should be noted that theantibody developed and used for immunohistochemical analysis would notdetect the TADG-12V truncated protein.

[0091] The results of the immunohistochemical staining are summarized inTable 3. 22 of 29 ovarian tumors showed positive staining of TADG-12,whereas normal ovarian surface epithelium showed no expression of theTADG-12 antigen. 8 of 10 serous adenocarcinomas, 8 of 8 mucinousadenocarcinomas, 1 of 2 clear cell carcinomas, and 4 of 6 endometroidcarcinomas showed positive staining. TABLE 3 Case Stage Histology GradeLN^(*) TADG12 Prognosis 1 Normal ovary 0− 2 Normal ovary 0− 3 Normalovary 0− 4 Mucinous B ND 0− Alive 5 Mucinous B ND 1+ Alive 6 1a SerousLMP G1 ND 1+ Alive 7 1a Mucinous LMP G1 ND 1+ Alive 8 1a Mucinous CA G1ND 1+ Alive 9 1a Mucinous CA G2 ND 1+ Alive 10 1a Endometrioid CA G1 ND0− Alive 11 1c Serous CA G1 N 1+ Alive 12 1c Mucinous CA G1 N 1+ Alive13 1c Mucinous CA G1 N 2+ Alive 14 1c Clear cell CA G2 N 0− Alive 15 1cClear cell CA G2 N 0− Alive 16 2c Serous CA G3 N 2+ Alive 17 3a MucinousCA G2 N 2+ Alive 18 3b Serous CA G1 ND 1+ Alive 19 3c Serous CA G1 N 0−Dead 20 3c Serous CA G3 P 1+ Alive 21 3c Serous CA G2 P 2+ Alive 22 3cSerous CA G1 P 2+ Unknown 23 3c Serous CA G3 ND 2+ Alive 24 3c Serous CAG2 N 0− Dead 25 3c Mucinous CA G1 P 2+ Dead 26 3c Mucinous CA G2 ND 1+Unknown 27 3c Mucinous CA G2 N 1+ Alive 28 3c Endometrioid CA G1 P 1+Dead 29 3c Endometrioid CA G2 N 0− Alive 30 3c Endometrioid CA G2 P 1+Dead 31 3c Endometrioid CA G3 P 1+ Alive 32 3c Clear Cell CA G3 P 2+Dead

EXAMPLE 14

[0092] Splicing Variant TADG-12D

[0093] The TADG-12V protein is generated from a splicing error thatoccurred downstream from the sentinel histidine (FIG. 9). The netoutcome of this splicing error is the inclusion of intron sequencedownstream from the histidine that has a stop codon 33 amino acidsdownstream.

[0094] A second splicing variant of TADG-12 was identified. This newsplicing error occurs downstream from the TADG-12V variant near thesentinel aspartic acid of the serine protease. It also results in theinclusion of intron sequence and a new stop codon. The result is avariant that includes unique amino acid sequence (TADG-12D, FIG. 10).TADG-12D is highly expressed in ovarian carcinoma, but not in normalovary (FIG. 11). TADG-12D is also expressed in breast and lung carcinoma(FIG. 12). The presence of unique peptide sequence in tumor cells offersa specific target and/or diagnostic marker for cancer treatment andmonitoring.

EXAMPLE 15

[0095] Peptide Ranking

[0096] For vaccine or immune stimulation, individual 9-mers to 11-mersof the TADG-12 protein were examined to rank the binding of individualpeptides to the top 8 haplotypes in the general population [Parker etal., (1994)]. The computer program used for this analysis can be foundon the internet. Table 4 shows the peptide ranking based upon thepredicted half-life of each peptide's binding to a particular HLAallele. A larger half-life indicates a stronger association with thatpeptide and the particular HLA molecule. The TADG-12 peptides thatstrongly bind to an HLA allele are putative immunogens, and are used toinnoculate an individual against TADG-12. TABLE 4 TADG-12 peptideranking HLA Type & Predicted SEQ Ranking Start PeptideDissociation_(1/2) ID No. HLA A0201 1 40 ILSLLPFEV 685.783 35 2 144AQLGFPSYV 545.316 36 3 225 LLSQWPWQA 63.342 37 4 252 WIITAAHCV 43.992 385 356 VLNHAAVPL 36.316 39 6 176 LLPDDKVTA 34.627 40 7 13 FSFRSLFGL31.661 41 8 151 YVSSDNLRV 27.995 42 9 436 RVTSFLDWI 21.502 43 10 234SLQFQGYHL 21.362 44 11 181 KVTALHHSV 21.300 45 12 183 TALHHSVYV 19.65846 13 411 RLWKLVGAT 18.494 47 14 60 LILALAIGL 18.476 48 15 227 SQWPWQASL17.977 49 16 301 RLGNDIALM 11.426 50 17 307 ALMKLAGPL 10.275 51 18 262DLYLPKSWT 9.837 52 19 416 LVGATSFGI 9.001 53 20 54 SLGIIALIL 8.759 54HLA A0205 1 218 IVGGNMSLL 47.600 55 2 60 LILALAIGL 35.700 48 3 35AVAAQILSL 28.000 56 4 307 ALMKLAGPL 21.000 51 5 271 IQVGLVSLL 19.040 576 397 CQGDSGGPL 16.800 58 7 227 SQWPWQASL 16.800 49 8 270 TIQVGLVSL14.000 59 9 56 GIIALILAL 14.000 60 10 110 RVGGQNAVL 14.000 61 11 181KVTALHHSV 12.000 45 12 151 YVSSDNLRV 12.000 42 13 356 VLNHAAVPL 11.90039 14 144 AQLGFPSYV 9.600 36 15 13 FSFRSLFGL 7.560 41 16 54 SLGIIALIL7.000 54 17 234 SLQFQGYHL 7.000 44 18 217 RIVGGNMSL 7.000 62 19 411RLWKLVGAT 6.000 47 20 252 WIITAAHCV 6.000 38 HLA A1 1 130 CSDDWKGHY37.500 63 2 8 AVEAPFSFR 9.000 64 3 328 NSEENFPDG 2.700 65 4 3 ENDPPAVEA2.500 66 5 98 DCKDGEDEY 2.500 67 6 346 ATEDGGDAS 2.250 68 7 360AAVPLISNK 2.000 69 8 153 SSDNLRVSS 1.500 70 9 182 VTALHHSVY 1.250 71 10143 CAQLGFPSY 1.000 72 11 259 CVYDLYLPK 1.000 73 12 369 ICNHRDVYG 1.00074 13 278 LLDNPAPSH 1.000 75 14 426 CAEVNKPGV 1.000 76 15 32 DADAVAAQI1.000 77 16 406 VCQERRLWK 1.000 78 17 329 SEENFPDGK 0.900 79 18 303GNDIALMKL 0.625 80 19 127 KTMCSDDWK 0.500 81 20 440 FLDWTHEQM 0.500 82HLA A24 1 433 VYIRVTSFL 280.000 83 2 263 LYLPKSWTI 90.000 84 3 169EFVSIDHLL 42.000 85 4 217 RIVGGNMSL 12.000 62 5 296 KYKPKRLGN 12.000 866 16 RSLFGLDDL 12.000 87 7 267 KSWTIQVGL 11.200 88 8 81 RSSFKCIEL 8.80089 9 375 VYGGIISPS 8.000 90 10 110 RVGGQNAVL 8.000 91 11 189 VYVREGCAS7.500 92 12 60 LILALAIGL 7.200 48 13 165 QFREEFVSI 7.200 93 14 271IQVGLVSLL 7.200 57 15 56 GIIALILAL 7.200 60 16 10 EAPFSFRSL 7.200 94 17307 ALMKLAGPL 7.200 51 18 407 CQERRLWKL 6.600 95 19 356 VLNHAAVPL 6.00039 20 381 SPSMLCAGY 6.000 96 HLA B7 1 375 VYGGIISPS 200.000 97 2 381SPSMLCAGY 80.000 98 3 362 VPLISNKIC 80.000 99 4 35 AVAAQILSL 60.000 56 5373 RDVYGGIIS 40.000 100 6 307 ALMKLAGPL 36.000 51 7 283 APSHLVEKI24.000 101 8 177 LPDDKVTAL 24.000 102 9 47 EVFSQSSSL 20.000 103 10 110RVGGQNAVL 20.000 91 11 218 IVGGNMSLL 20.000 55 12 36 VAAQTLSLL 12.000104 13 255 TAAHCVYDL 12.000 105 14 10 EAPFSFRSL 12.000 94 15 138YANVACAQL 12.000 106 16 195 CASGHVVTL 12.000 107 17 215 SSRIVGGNM 10.000108 18 298 KPKRLGNDI 8.000 109 19 313 GPLTFNEMI 8.000 110 20 108CVRVGGQNA 5.000 111 HLA B8 1 294 HSKYKPKRL 80.000 112 2 373 RDVYGGIIS16.000 100 3 177 LPDDKVTAL 4.800 102 4 265 LPKSWTIQV 2.400 113 5 88ELITRCDGV 2.400 114 6 298 KPKRLGNDI 2.000 109 7 81 RSSFKCIEL 2.000 89 8375 VYGGIISPS 2.000 97 9 79 RCRSSFKCI 2.000 115 10 10 EAPFSFRSL 1.600 9411 215 SSRIVGGNM 1.000 108 12 36 VAAQILSLL 0.800 104 13 255 TAAHCVYDL0.800 116 14 381 SPSMLCAGY 0.800 98 15 195 CASGHVVTL 0.800 107 16 362VPLISNKIC 0.800 99 17 138 YANVACAQL 0.800 106 18 207 ACGHRRGYS 0.400 11719 154 SDNLRVSSL 0.400 118 20 47 EVFSQSSSL 0.400 103 HLA B2702 1 300KRLGNDIAL 180.000 119 2 435 TRVTSFLDW 100.000 120 3 376 YGGIISPSM100.000 121 4 410 RRLWKLVGA 60.000 122 5 210 HRRGYSSRI 60.000 123 6 227SQWPWQASL 30.000 49 7 109 VRVGGQNAV 20.000 124 8 191 VREGCASGH 20.000125 9 78 YRCRSSFKC 20.000 126 10 113 GQNAVLQVF 20.000 127 11 91TRCDGVSDC 20.000 128 12 38 AQILSLLPF 20.000 129 13 211 RRGYSSRIV 18.000130 14 216 SRIVGGNMS 10.000 131 15 118 LQVFTAASW 10.000 132 16 370CNHRDVYGG 10.000 133 17 393 GVDSCQGDS 10.000 134 18 235 LQFQGYHLC 10.000135 19 271 IQVGLVSLL 6.000 57 20 408 CQERRLWKL 6.000 95 HLA B4403 1 427AEVNKPGVY 90.000 136 2 162 LEGQFREEF 40.000 137 3 9 VEAPFSFRS 24.000 1384 318 NEMIQPVCL 12.000 139 5 256 AAHCVYDLY 9.000 140 6 98 DCKDGEDEY9.000 67 7 46 FEVFSQSSS 8.000 141 8 38 AQILSLLPF 7.500 129 9 64LAIGLGIHF 7.500 142 10 192 REGCASGHV 6.000 143 11 330 EENFPDGKV 6.000144 12 182 VTALHHSVY 6.000 145 13 408 QERRLWKLV 6.000 146 14 206TACGHRRGY 4.500 147 15 5 DPPAVEAPF 4.500 148 16 261 YDLYLPKSW 4.500 14917 33 ADAVAAQIL 4.500 150 18 168 EEFVSIDHL 4.000 151 19 304 NDIALMKLA3.750 152 20 104 DEYRCVRVG 3.600 153

[0097] The following references were cited herein.

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[0112] 15. Maniatis, T., Fritsch, E. F. & Sambrook, J. MolecularCloning, p. 309-361 Cold Spring Harbor Laboratory, New York, 1982.

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[0120] Any patents or publications mentioned in this specification areindicative of the levels of those skilled in the art to which theinvention pertains. These patents and publications are hereinincorporated by reference to the same extent as if each individualpublication was specifically and individually indicated to b eincorporated by reference.

[0121] One skilled in the art will readily appreciate that the presentinvention is well adapted to carry out the objects and obtain the endsand advantages mentioned, as well as those inherent therein. The presentexamples along with the methods, procedures, treatments, molecules, andspecific compounds described herein are presently representative ofpreferred embodiments, are exemplary, and are not intended aslimitations on the scope of the invention. Changes therein and otheruses will occur to those skilled in the art which are encompassed withinthe spirit of the invention as defined by the scope of the claims.

1 158 1 2413 DNA Homo sapiens CDS entire cDNA sequence of TADG-12 gene 1cgggaaaggg ctgtgtttat gggaagccag taacactgtg gcctactatc 50 tcttccgtggtgccatctac atttttggga ctcgggaatt atgaggtaga 100 ggtggaggcg gagccggatgtcagaggtcc tgaaatagtc accatggggg 150 aaaatgatcc gcctgctgtt gaagcccccttctcattccg atcgcttttt 200 ggccttgatg atttgaaaat aagtcctgtt gcaccagatgcagatgctgt 250 tgctgcacag atcctgtcac tgctgccatt tgaagttttt tcccaatcat300 cgtcattggg gatcattgca ttgatattag cactggccat tggtctgggc 350atccacttcg actgctcagg gaagtacaga tgtcgctcat cctttaagtg 400 tatcgagctgataactcgat gtgacggagt ctcggattgc aaagacgggg 450 aggacgagta ccgctgtgtccgggtgggtg gtcagaatgc cgtgctccag 500 gtgttcacag ctgcttcgtg gaagaccatgtgctccgatg actggaaggg 550 tcactacgca aatgttgcct gtgcccaact gggtttcccaagctatgtga 600 gttcagataa cctcagagtg agctcgctgg aggggcagtt ccgggaggag650 tttgtgtcca tcgatcacct cttgccagat gacaaggtga ctgcattaca 700ccactcagta tatgtgaggg agggatgtgc ctctggccac gtggttacct 750 tgcagtgcacagcctgtggt catagaaggg gctacagctc acgcatcgtg 800 ggtggaaaca tgtccttgctctcgcagtgg ccctggcagg ccagccttca 850 gttccagggc taccacctgt gcgggggctctgtcatcacg cccctgtgga 900 tcatcactgc tgcacactgt gtttatgact tgtacctccccaagtcatgg 950 accatccagg tgggtctagt ttccctgttg gacaatccag ccccatccca1000 cttggtggag aagattgtct accacagcaa gtacaagcca aagaggctgg 1050gcaatgacat cgcccttatg aagctggccg ggccactcac gttcaatgaa 1100 atgatccagcctgtgtgcct gcccaactct gaagagaact tccccgatgg 1150 aaaagtgtgc tggacgtcaggatggggggc cacagaggat ggaggtgacg 1200 cctcccctgt cctgaaccac gcggccgtccctttgatttc caacaagatc 1250 tgcaaccaca gggacgtgta cggtggcatc atctccccctccatgctctg 1300 cgcgggctac ctgacgggtg gcgtgaacag ctgccagggg gacagcgggg1350 ggcccctggt gtgtcaagag aggaggctgt ggaagttagt gggagcgacc 1400agctttggca tcggctgcgc agaggtgaac aagcctgggg tgtacacccg 1450 tgtcacctccttcctggact ggatccacga gcagatggag agagacctaa 1500 aaacctgaag aggaaggggacaagtagcca cctgagttcc tgaggtgatg 1550 aagacagccc gatcctcccc tggactcccgtgtaggaacc tgcacacgag 1600 cagacaccct tggagctctg agttccggca ccagtagcgggcccgaaaga 1650 ggcacccttc catctgattc cagcacaacc ttcaagctgc tttttgtttt1700 ttgttttttt gaggtggagt ctcgctctgt tgcccaggct ggagtgcagt 1750ggcgaaatac cctgctcact gcagcctccg cttccctggt tcaagcgatt 1800 ctcttgcctcagcttcccca gtagctggga ccacaggtgc ccgccaccac 1850 acccaactaa tttttgtatttttagtagag acagggtttc accatgttgg 1900 ccaggctgct ctcaaacccc tgacctcaaatgatgtgcct gcttcagcct 1950 cccacagtgc tgggattaca ggcatgggcc accacgcctagcctcacgct 2000 cctttctgat cttcactaag aacaaaagaa gcagcaactt gcaagggcgg2050 cctttcccac tggtccatct ggttttctct ccagggtctt gcaaaattcc 2100tgacgagata agcagttatg tgacctcacg tgcaaagcca ccaacagcca 2150 ctcagaaaagacgcaccagc ccagaagtgc agaactgcag tcactgcacg 2200 ttttcatctt tagggaccagaaccaaaccc accctttcta cttccaagac 2250 ttattttcac atgtggggag gttaatctaggaatgactcg tttaaggcct 2300 attttcatga tttctttgta gcatttggtg cttgacgtattattgtcctt 2350 tgattccaaa taatatgttt ccttccctca aaaaaaaaaa aaaaaaaaaa2400 aaaaaaaaaa aaa 2413 2 454 PRT Homo sapiens complete amino acidsequence of TADG-12 protein 2 Met Gly Glu Asn Asp Pro Pro Ala Val GluAla Pro Phe Ser Phe 5 10 15 Arg Ser Leu Phe Gly Leu Asp Asp Leu Lys IleSer Pro Val Ala 20 25 30 Pro Asp Ala Asp Ala Val Ala Ala Gln Ile Leu SerLeu Leu Pro 35 40 45 Phe Glu Val Phe Ser Gln Ser Ser Ser Leu Gly Ile IleAla Leu 50 55 60 Ile Leu Ala Leu Ala Ile Gly Leu Gly Ile His Phe Asp CysSer 65 70 75 Gly Lys Tyr Arg Cys Arg Ser Ser Phe Lys Cys Ile Glu Leu Ile80 85 90 Thr Arg Cys Asp Gly Val Ser Asp Cys Lys Asp Gly Glu Asp Glu 95100 105 Tyr Arg Cys Val Arg Val Gly Gly Gln Asn Ala Val Leu Gln Val 110115 120 Phe Thr Ala Ala Ser Trp Lys Thr Met Cys Ser Asp Asp Trp Lys 125130 135 Gly His Tyr Ala Asn Val Ala Cys Ala Gln Leu Gly Phe Pro Ser 140145 150 Tyr Val Ser Ser Asp Asn Leu Arg Val Ser Ser Leu Glu Gly Gln 155160 165 Phe Arg Glu Glu Phe Val Ser Ile Asp His Leu Leu Pro Asp Asp 170175 180 Lys Val Thr Ala Leu His His Ser Val Tyr Val Arg Glu Gly Cys 185190 195 Ala Ser Gly His Val Val Thr Leu Gln Cys Thr Ala Cys Gly His 200205 210 Arg Arg Gly Tyr Ser Ser Arg Ile Val Gly Gly Asn Met Ser Leu 215220 225 Leu Ser Gln Trp Pro Trp Gln Ala Ser Leu Gln Phe Gln Gly Tyr 230235 240 His Leu Cys Gly Gly Ser Val Ile Thr Pro Leu Trp Ile Ile Thr 245250 255 Ala Ala His Cys Val Tyr Asp Leu Tyr Leu Pro Lys Ser Trp Thr 260265 270 Ile Gln Val Gly Leu Val Ser Leu Leu Asp Asn Pro Ala Pro Ser 275280 285 His Leu Val Glu Lys Ile Val Tyr His Ser Lys Tyr Lys Pro Lys 290295 300 Arg Leu Gly Asn Asp Ile Ala Leu Met Lys Leu Ala Gly Pro Leu 305310 315 Thr Phe Asn Glu Met Ile Gln Pro Val Cys Leu Pro Asn Ser Glu 320325 330 Glu Asn Phe Pro Asp Gly Lys Val Cys Trp Thr Ser Gly Trp Gly 335340 345 Ala Thr Glu Asp Gly Gly Asp Ala Ser Pro Val Leu Asn His Ala 350355 360 Ala Val Pro Leu Ile Ser Asn Lys Ile Cys Asn His Arg Asp Val 365370 375 Tyr Gly Gly Ile Ile Ser Pro Ser Met Leu Cys Ala Gly Tyr Leu 380385 390 Thr Gly Gly Val Asp Ser Cys Gln Gly Asp Ser Gly Gly Pro Leu 395400 405 Val Cys Gln Glu Arg Arg Leu Trp Lys Leu Val Gly Ala Thr Ser 410415 420 Phe Gly Ile Gly Cys Ala Glu Val Asn Lys Pro Gly Val Tyr Thr 425430 435 Arg Val Thr Ser Phe Leu Asp Trp Ile His Glu Gln Met Glu Arg 440445 450 Asp Leu Lys Thr 3 2544 DNA Homo sapiens CDS entire cDNA sequenceof TADG-12V 3 cgggaaaggg ctgtgtttat gggaagccag taacactgtg gcctactatc 50tcttccgtgg tgccatctac atttttggga ctcgggaatt atgaggtaga 100 ggtggaggcggagccggatg tcagaggtcc tgaaatagtc accatggggg 150 aaaatgatcc gcctgctgttgaagccccct tctcattccg atcgcttttt 200 ggccttgatg atttgaaaat aagtcctgttgcaccagatg cagatgctgt 250 tgctgcacag atcctgtcac tgctgccatt tgaagttttttcccaatcat 300 cgtcattggg gatcattgca ttgatattag cactggccat tggtctgggc350 atccacttcg actgctcagg gaagtacaga tgtcgctcat cctttaagtg 400tatcgagctg ataactcgat gtgacggagt ctcggattgc aaagacgggg 450 aggacgagtaccgctgtgtc cgggtgggtg gtcagaatgc cgtgctccag 500 gtgttcacag ctgcttcgtggaagaccatg tgctccgatg actggaaggg 550 tcactacgca aatgttgcct gtgcccaactgggtttccca agctatgtaa 600 gttcagataa cctcagagtg agctcgctgg aggggcagttccgggaggag 650 tttgtgtcca tcgatcacct cttgccagat gacaaggtga ctgcattaca700 ccactcagta tatgtgaggg agggatgtgc ctctggccac gtggttacct 750tgcagtgcac agcctgtggt catagaaggg gctacagctc acgcatcgtg 800 ggtggaaacatgtccttgct ctcgcagtgg ccctggcagg ccagccttca 850 gttccagggc taccacctgtgcgggggctc tgtcatcacg cccctgtgga 900 tcatcactgc tgcacactgt gtttatgagattgtagctcc tagagaaagg 950 gcagacagaa gaggaaggaa gctcctgtgc tggaggaaacccacaaaaat 1000 gaaaggacct agaccttccc atagctaatt ccagtggacc atgttatggc1050 agatacaggc ttgtacctcc ccaagtcatg gaccatccag gtgggtctag 1100tttccctgtt ggacaatcca gccccatccc acttggtgga gaagattgtc 1150 taccacagcaagtacaagcc aaagaggctg ggcaatgaca tcgcccttat 1200 gaagctggcc gggccactcacgttcaatga aatgatccag cctgtgtgcc 1250 tgcccaactc tgaagagaac ttccccgatggaaaagtgtg ctggacgtca 1300 ggatgggggg ccacagagga tggaggtgac gcctcccctgtcctgaacca 1350 cgcggccgtc cctttgattt ccaacaagat ctgcaaccac agggacgtgt1400 acggtggcat catctccccc tccatgctct gcgcgggcta cctgacgggt 1450ggcgtggaca gctgccaggg ggacagcggg gggcccctgg tgtgtcaaga 1500 gaggaggctgtggaagttag tgggagcgac cagctttggc atcggctgcg 1550 cagaggtgaa caagcctggggtgtacaccc gtgtcacctc cttcctggac 1600 tggatccacg agcagatgga gagagacctaaaaacctgaa gaggaagggg 1650 acaagtagcc acctgagttc ctgaggtgat gaagacagcccgatcctccc 1700 ctggactccc gtgtaggaac ctgcacacga gcagacaccc ttggagctct1750 gagttccggc accagtagcg ggcccgaaag aggcaccctt ccatctgatt 1800ccagcacaac cttcaagctg ctttttgttt tttgtttttt tgaggtggag 1850 tctcgctctgttgcccaggc tggagtgcag tggcgaaata ccctgctcac 1900 tgcagcctcc gcttccctggttcaagcgat tctcttgcct cagcttcccc 1950 agtagctggg accacaggtg cccgccaccacacccaacta atttttgtat 2000 ttttagtaga gacagggttt caccatgttg gccaggctgctctcaaaccc 2050 ctgacctcaa atgatgtgcc tgcttcagcc tcccacagtg ctgggattac2100 aggcatgggc caccacgcct agcctcacgc tcctttctga tcttcactaa 2150gaacaaaaga agcagcaact tgcaagggcg gcctttccca ctggtccatc 2200 tggttttctctccagggtct tgcaaaattc ctgacgagat aagcagttat 2250 gtgacctcac gtgcaaagccaccaacagcc actcagaaaa gacgcaccag 2300 cccagaagtg cagaactgca gtcactgcacgttttcatct ttagggacca 2350 gaaccaaacc caccctttct acttccaaga cttattttcacatgtgggga 2400 ggttaatcta ggaatgactc gtttaaggcc tattttcatg atttctttgt2450 agcatttggt gcttgacgta ttattgtcct ttgattccaa ataatatgtt 2500tccttccctc aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaa 2544 4 294 PRT Homosapiens complete amino acid sequence of TADG-12V 4 Met Gly Glu Asn AspPro Pro Ala Val Glu Ala Pro Phe Ser Phe 5 10 15 Arg Ser Leu Phe Gly LeuAsp Asp Leu Lys Ile Ser Pro Val Ala 20 25 30 Pro Asp Ala Asp Ala Val AlaAla Gln Ile Leu Ser Leu Leu Pro 35 40 45 Phe Glu Val Phe Ser Gln Ser SerSer Leu Gly Ile Ile Ala Leu 50 55 60 Ile Leu Ala Leu Ala Ile Gly Leu GlyIle His Phe Asp Cys Ser 65 70 75 Gly Lys Tyr Arg Cys Arg Ser Ser Phe LysCys Ile Glu Leu Ile 80 85 90 Thr Arg Cys Asp Gly Val Ser Asp Cys Lys AspGly Glu Asp Glu 95 100 105 Tyr Arg Cys Val Arg Val Gly Gly Gln Asn AlaVal Leu Gln Val 110 115 120 Phe Thr Ala Ala Ser Trp Lys Thr Met Cys SerAsp Asp Trp Lys 125 130 135 Gly His Tyr Ala Asn Val Ala Cys Ala Gln LeuGly Phe Pro Ser 140 145 150 Tyr Val Ser Ser Asp Asn Leu Arg Val Ser SerLeu Glu Gly Gln 155 160 165 Phe Arg Glu Glu Phe Val Ser Ile Asp His LeuLeu Pro Asp Asp 170 175 180 Lys Val Thr Ala Leu His His Ser Val Tyr ValArg Glu Gly Cys 185 190 195 Ala Ser Gly His Val Val Thr Leu Gln Cys ThrAla Cys Gly His 200 205 210 Arg Arg Gly Tyr Ser Ser Arg Ile Val Gly GlyAsn Met Ser Leu 215 220 225 Leu Ser Gln Trp Pro Trp Gln Ala Ser Leu GlnPhe Gln Gly Tyr 230 235 240 His Leu Cys Gly Gly Ser Val Ile Thr Pro LeuTrp Ile Ile Thr 245 250 255 Ala Ala His Cys Val Tyr Glu Ile Val Ala ProArg Glu Arg Ala 260 265 270 Asp Arg Arg Gly Arg Lys Leu Leu Cys Trp ArgLys Pro Thr Lys 275 280 285 Met Lys Gly Pro Arg Pro Ser His Ser 290 5174 DNA Artificial sequence nucleotide sequence of the subclonecontaining the 180 bp band from the PCR product for TADG-12 5 tgggtggtgacggcggcgca ctgtgtttat gacttgtacc tccccaagtc 50 atggaccatc caggtgggtctagtttccct gttggacaat ccagccccat 100 cccacttggt ggagaagatt gtctaccacagcaagtacaa gccaaagagg 150 ctgggcaacg acatcgccct ccta 174 6 58 PRTArtificial sequence deduced amino acid sequence of the 180 bp band fromthe PCR product for TADG-12 6 Trp Val Val Thr Ala Ala His Cys Val TyrAsp Leu Tyr Leu Pro 5 10 15 Lys Ser Trp Thr Ile Gln Val Gly Leu Val SerLeu Leu Asp Asn 20 25 30 Pro Ala Pro Ser His Leu Val Glu Lys Ile Val TyrHis Ser Lys 35 40 45 Tyr Lys Pro Lys Arg Leu Gly Asn Asp Ile Ala Leu Leu50 55 7 328 DNA Artificial sequence nucleotide sequence of the subclonecontaining the 300 bp band from the PCR product for TADG-12 variant,which contains an additional insert of 133 bases 7 gggtggtgac ggcggcgcactgtgtttatg agattgtagc tcctagagaa 50 agggcagaca gaagaggaag gaagctcctgtgctggagga aacccacaaa 100 aatgaaagga cctagacctt cccatagcta attccagtggaccatgttat 150 ggcagataca ggcttgtacc tccccaagtc atggaccatc caggtgggtc200 tagtttccct gttggacaat ccagccccat cccacttggt ggagaagatt 250gtctaccaca gcaagtacaa gccaaagagg ctgggcaacg acatcgccct 300 cctaatcactagtgcggccg cctgcagg 328 8 42 PRT Artificial sequence deduced amino acidsequence of the 300 bp band from the PCR product for TADG-12 variant,which is a truncated form of TADG-12 8 Val Val Thr Ala Ala His Cys ValTyr Glu Ile Val Ala Pro Arg 5 10 15 Glu Arg Ala Asp Arg Arg Gly Arg LysLeu Leu Cys Trp Arg Lys 20 25 30 Pro Thr Lys Met Lys Gly Pro Arg Pro SerHis Ser 35 40 9 34 PRT Homo sapiens DOMAIN LDLR-A domain of thecomplement subunit C8 (Compc8) 9 Cys Glu Gly Phe Val Cys Ala Gln Thr GlyArg Cys Val Asn Arg 5 10 15 Arg Leu Leu Cys Asn Gly Asp Asn Asp Cys GlyAsp Gln Ser Asp 20 25 30 Glu Ala Asn Cys 10 34 PRT Homo sapiens DOMAINLDLR-A domain of the serine protease matriptase (Matr) 10 Cys Pro GlyGln Phe Thr Cys Arg Thr Gly Arg Cys Ile Arg Lys 5 10 15 Glu Leu Arg CysAsp Gly Trp Ala Asp Cys Thr Asp His Ser Asp 20 25 30 Glu Leu Asn Cys 1137 PRT Homo sapiens DOMAIN LDLR-A domain of the glycoprotein GP300(Gp300-1) 11 Cys Gln Gln Gly Tyr Phe Lys Cys Gln Ser Glu Gly Gln Cys Ile5 10 15 Pro Ser Ser Trp Val Cys Asp Gln Asp Gln Asp Cys Asp Asp Gly 2025 30 Ser Asp Glu Arg Gln Asp Cys 35 12 35 PRT Homo sapiens DOMAINLDLR-A domain of the glycoprotein GP300 (Gp300-2) 12 Cys Ser Ser His GlnIle Thr Cys Ser Asn Gly Gln Cys Ile Pro 5 10 15 Ser Glu Tyr Arg Cys AspHis Val Arg Asp Cys Pro Asp Gly Ala 20 25 30 Asp Glu Asn Asp Cys 35 1335 PRT Homo sapiens DOMAIN 74...108 LDLR-A domain of TADG-12 13 Cys SerGly Lys Tyr Arg Cys Arg Ser Ser Phe Lys Cys Ile Glu 5 10 15 Leu Ile ThrArg Cys Asp Gly Val Ser Asp Cys Lys Asp Gly Glu 20 25 30 Asp Glu Tyr ArgCys 35 14 36 PRT Homo sapiens DOMAIN LDLR-A domain of the serineprotease TMPRSS2 Tmprss2 14 Cys Ser Asn Ser Gly Ile Glu Cys Asp Ser SerGly Thr Cys Ile 5 10 15 Asn Pro Ser Asn Trp Cys Asp Gly Val Ser His CysPro Gly Gly 20 25 30 Glu Asp Glu Asn Arg Cys 35 15 101 PRT Bos taurusDOMAIN SRCR domain of bovine enterokinase (BovEntk) 15 Val Arg Leu ValGly Gly Ser Gly Pro His Glu Gly Arg Val Glu 5 10 15 Ile Phe His Glu GlyGln Trp Gly Thr Val Cys Asp Asp Arg Trp 20 25 30 Glu Leu Arg Gly Gly LeuVal Val Cys Arg Ser Leu Gly Tyr Lys 35 40 45 Gly Val Gln Ser Val His LysArg Ala Tyr Phe Gly Lys Gly Thr 50 55 60 Gly Pro Ile Trp Leu Asn Glu ValPhe Cys Phe Gly Lys Glu Ser 65 70 75 Ser Ile Glu Glu Cys Arg Ile Arg GlnTrp Gly Val Arg Ala Cys 80 85 90 Ser His Asp Glu Asp Ala Gly Val Thr CysThr 95 100 16 101 PRT Homo sapiens DOMAIN SRCR domain of humanmacrophage scavenger receptor (MacSR) 16 Val Arg Leu Val Gly Gly Ser GlyPro His Glu Gly Arg Val Glu 5 10 15 Ile Leu His Ser Gly Gln Trp Gly ThrIle Cys Asp Asp Arg Trp 20 25 30 Glu Val Arg Val Gly Gln Val Val Cys ArgSer Leu Gly Tyr Pro 35 40 45 Gly Val Gln Ala Val His Lys Ala Ala His PheGly Gln Gly Thr 50 55 60 Gly Pro Ile Trp Leu Asn Glu Val Phe Cys Phe GlyArg Glu Ser 65 70 75 Ser Ile Glu Glu Cys Lys Ile Arg Gln Trp Gly Thr ArgAla Cys 80 85 90 Ser His Ser Glu Asp Ala Gly Val Thr Cys Thr 95 100 1798 PRT Homo sapiens DOMAIN 109...206 SRCR domain of TADG-12 (TADG12) 17Val Arg Val Gly Gly Gln Asn Ala Val Leu Gln Val Phe Thr Ala 5 10 15 AlaSer Trp Lys Thr Met Cys Ser Asp Asp Trp Lys Gly His Tyr 20 25 30 Ala AsnVal Ala Cys Ala Gln Leu Gly Phe Pro Ser Tyr Val Ser 35 40 45 Ser Asp AsnLeu Arg Val Ser Ser Leu Glu Gly Gln Phe Arg Glu 50 55 60 Glu Phe Val SerIle Asp His Leu Leu Pro Asp Asp Lys Val Thr 65 70 75 Ala Leu His His SerVal Tyr Val Arg Glu Gly Cys Ala Ser Gly 80 85 90 His Val Val Thr Leu GlnCys Thr 95 18 94 PRT Homo sapiens DOMAIN SRCR domain of the serineprotease TMPRSS2 (Tmprss2) 18 Val Arg Leu Tyr Gly Pro Asn Phe Ile LeuGln Met Tyr Ser Ser 5 10 15 Gln Arg Lys Ser Trp His Pro Val Cys Gln AspAsp Trp Asn Glu 20 25 30 Asn Tyr Gly Arg Ala Ala Cys Arg Asp Met Gly TyrLys Asn Asn 35 40 45 Phe Tyr Ser Ser Gln Gly Ile Val Asp Asp Ser Gly SerThr Ser 50 55 60 Phe Met Lys Leu Asn Thr Ser Ala Gly Asn Val Asp Ile TyrLys 65 70 75 Lys Leu Tyr His Ser Asp Ala Cys Ser Ser Lys Ala Val Val Ser80 85 90 Leu Arg Cys Leu 19 90 PRT Homo sapiens DOMAIN SRCR domain ofhuman enterokinase (HumEntk) 19 Val Arg Phe Phe Asn Gly Thr Thr Asn AsnAsn Gly Leu Val Arg 5 10 15 Phe Arg Ile Gln Ser Ile Trp His Thr Ala CysAla Glu Asn Trp 20 25 30 Thr Thr Gln Ile Ser Asn Asp Val Cys Gln Leu LeuGly Leu Gly 35 40 45 Ser Gly Asn Ser Ser Lys Pro Ile Phe Ser Thr Asp GlyGly Pro 50 55 60 Phe Val Lys Leu Asn Thr Ala Pro Asp Gly His Leu Ile LeuThr 65 70 75 Pro Ser Gln Gln Cys Leu Gln Asp Ser Leu Ile Arg Leu Gln Cys80 85 90 20 149 PRT Homo sapiens DOMAIN protease domain of protease M(ProM) 20 Leu Trp Val Leu Thr Ala Ala His Cys Lys Lys Pro Asn Leu Gln 510 15 Val Phe Leu Gly Lys His Asn Leu Arg Gln Arg Glu Ser Ser Gln 20 2530 Glu Gln Ser Ser Val Val Arg Ala Val Ile His Pro Asp Tyr Asp 35 40 45Ala Ala Ser His Asp Gln Asp Ile Met Leu Leu Arg Leu Ala Arg 50 55 60 ProAla Lys Leu Ser Glu Leu Ile Gln Pro Leu Pro Leu Glu Arg 65 70 75 Asp CysSer Ala Asn Thr Thr Ser Cys His Ile Leu Gly Trp Gly 80 85 90 Lys Thr AlaAsp Gly Asp Phe Pro Asp Thr Ile Gln Cys Ala Tyr 95 100 105 Ile His LeuVal Ser Arg Glu Glu Cys Glu His Ala Tyr Pro Gly 110 115 120 Gln Ile ThrGln Asn Met Leu Cys Ala Gly Asp Glu Lys Tyr Gly 125 130 135 Lys Asp SerCys Gln Gly Asp Ser Gly Gly Pro Leu Val Cys 140 145 21 151 PRT Homosapiens DOMAIN protease domain of trypsinogen I (Try1) 21 Gln Trp ValVal Ser Ala Gly His Cys Tyr Lys Ser Arg Ile Gln 5 10 15 Val Arg Leu GlyGlu His Asn Ile Glu Val Leu Glu Gly Asn Glu 20 25 30 Gln Phe Ile Asn AlaAla Lys Ile Ile Arg His Pro Gln Tyr Asp 35 40 45 Arg Lys Thr Leu Asn AsnAsp Ile Met Leu Ile Lys Leu Ser Ser 50 55 60 Arg Ala Val Ile Asn Ala ArgVal Ser Thr Ile Ser Leu Pro Thr 65 70 75 Ala Pro Pro Ala Thr Gly Thr LysCys Leu Ile Ser Gly Trp Gly 80 85 90 Asn Thr Ala Ser Ser Gly Ala Asp TyrPro Asp Glu Leu Gln Cys 95 100 105 Leu Asp Ala Pro Val Leu Ser Gln AlaLys Cys Glu Ala Ser Tyr 110 115 120 Pro Gly Lys Ile Thr Ser Asn Met PheCys Val Gly Phe Leu Glu 125 130 135 Gly Gly Lys Asp Ser Cys Gln Gly AspSer Gly Gly Pro Val Val 140 145 150 Cys 22 158 PRT Homo sapiens DOMAINprotease domain of plasma kallikrein (Kal) 22 Gln Trp Val Leu Thr AlaAla His Cys Phe Asp Gly Leu Pro Leu 5 10 15 Gln Asp Val Trp Arg Ile TyrSer Gly Ile Leu Asn Leu Ser Asp 20 25 30 Ile Thr Lys Asp Thr Pro Phe SerGln Ile Lys Glu Ile Ile Ile 35 40 45 His Gln Asn Tyr Lys Val Ser Glu GlyAsn His Asp Ile Ala Leu 50 55 60 Ile Lys Leu Gln Ala Pro Leu Asn Tyr ThrGlu Phe Gln Lys Pro 65 70 75 Ile Cys Leu Pro Ser Lys Gly Asp Thr Ser ThrIle Tyr Thr Asn 80 85 90 Cys Trp Val Thr Gly Trp Gly Phe Ser Lys Glu LysGly Glu Ile 95 100 105 Gln Asn Ile Leu Gln Lys Val Asn Ile Pro Leu ValThr Asn Glu 110 115 120 Glu Cys Gln Lys Arg Tyr Gln Asp Tyr Lys Ile ThrGln Arg Met 125 130 135 Val Cys Ala Gly Tyr Lys Glu Gly Gly Lys Asp AlaCys Lys Gly 140 145 150 Asp Ser Gly Gly Pro Leu Val Cys 155 23 157 PRTHomo sapiens DOMAIN protease domain of TADG-12 (TADG12) 23 24 159 PRTHomo sapiens DOMAIN protease domain of TMPRSS2 (Tmprss2) 24 25 164 PRTHomo sapiens DOMAIN protease domain of Hepsin (Heps) 25 26 23 DNAArtificial sequence primer_bind 6, 9, 12, 15, 18 forward redundantprimer for the consensus sequences of amino acids surrounding thecatalytic triad for serine proteases, n = inosine 26 tgggtngtnacngcngcnca ytg 23 27 20 DNA Artificial sequence primer_bind 3, 6, 9, 12,15, 18 reverse redundant primer for the consensus sequences of aminoacids surrounding the catalytic triad for serine proteases, n = inosine27 arnarngcna tntcnttncc 20 28 20 DNA Artificial sequence primer_bindforward oligonucleotide primer for TADG-12 used for quantitative PCR 28gaaacatgtc cttgctctcg 20 29 20 DNA Artificial sequence primer_bindreverse oligonucleotide primer for TADG-12 used for quantitative PCR 29actaacttcc acagcctcct 20 30 20 DNA Artificial sequence primer_bindforward oligonucleotide primer for TADG-12 variant (TADG-12V) used forquantitative PCR 30 tccaggtggg tctagtttcc 20 31 20 DNA Artificialsequence primer_bind reverse oligonucleotide primer for TADG-12 variant(TADG-12V) used for quantitative PCR 31 ctctttggct tgtacttgct 20 32 20DNA Artificial sequence primer_bind forward oligonucleotide primer for(-tubulin used as an internal control for quantitative PCR 32 cgcatcaacgtgtactacaa 20 33 20 DNA Artificial sequence primer_bind reverseoligonucleotide primer for (-tubulin used as an internal control forquantitative PCR 33 tacgagctgg tggactgaga 20 34 12 PRT Artificialsequence a poly-lysine linked multiple antigen peptide derived from theTADG-12 carboxy-terminal protein sequence, present in full lengthTADG-12, but not in TADG-12V 34 Trp Ile His Glu Gln Met Glu Arg Asp LeuLys Thr 5 10 35 9 PRT Homo sapiens 40...48 TADG-12 peptide 35 Ile LeuSer Leu Leu Pro Phe Glu Val 5 36 9 PRT Homo sapiens 144...152 TADG-12peptide 36 Ala Gln Leu Gly Phe Pro Ser Tyr Val 5 37 9 PRT Homo sapiens225...233 TADG-12 peptide 37 Leu Leu Ser Gln Trp Pro Trp Gln Ala 5 38 9PRT Homo sapiens 252...260 TADG-12 peptide 38 Trp Ile Ile Thr Ala AlaHis Cys Val 5 39 9 PRT Homo sapiens 356...364 TADG-12 peptide 39 Val LeuAsn His Ala Ala Val Pro Leu 5 40 9 PRT Homo sapiens 176...184 TADG-12peptide 40 Leu Leu Pro Asp Asp Lys Val Thr Ala 5 41 9 PRT Homo sapiens13...21 TADG-12 peptide 41 Phe Ser Phe Arg Ser Leu Phe Gly Leu 5 42 9PRT Homo sapiens 151...159 TADG-12 peptide 42 Tyr Val Ser Ser Asp AsnLeu Arg Val 5 43 9 PRT Homo sapiens 436...444 TADG-12 peptide 43 Arg ValThr Ser Phe Leu Asp Trp Ile 5 44 9 PRT Homo sapiens 234...242 TADG-12peptide 44 Ser Leu Gln Phe Gln Gly Tyr His Leu 5 45 9 PRT Homo sapiens181...189 TADG-12 peptide 45 Lys Val Thr Ala Leu His His Ser Val 5 46 9PRT Homo sapiens 183...191 TADG-12 peptide 46 Thr Ala Leu His His SerVal Tyr Val 5 47 9 PRT Homo sapiens 411...419 TADG-12 peptide 47 Arg LeuTrp Lys Leu Val Gly Ala Thr 5 48 9 PRT Homo sapiens 60...68 TADG-12peptide 48 Leu Ile Leu Ala Leu Ala Ile Gly Leu 5 49 9 PRT Homo sapiens227...235 TADG-12 peptide 49 Ser Gln Trp Pro Trp Gln Ala Ser Leu 5 50 9PRT Homo sapiens 301...309 TADG-12 peptide 50 Arg Leu Gly Asn Asp IleAla Leu Met 5 51 9 PRT Homo sapiens 307...315 TADG-12 peptide 51 Ala LeuMet Lys Leu Ala Gly Pro Leu 5 52 9 PRT Homo sapiens 262...270 TADG-12peptide 52 Asp Leu Tyr Leu Pro Lys Ser Trp Thr 5 53 9 PRT Homo sapiens416...424 TADG-12 peptide 53 Leu Val Gly Ala Thr Ser Phe Gly Ile 5 54 9PRT Homo sapiens 54...62 TADG-12 peptide 54 Ser Leu Gly Ile Ile Ala LeuIle Leu 5 55 9 PRT Homo sapiens 218...226 TADG-12 peptide 55 Ile Val GlyGly Asn Met Ser Leu Leu 5 56 9 PRT Homo sapiens 35...43 TADG-12 peptide56 Ala Val Ala Ala Gln Ile Leu Ser Leu 5 57 9 PRT Homo sapiens 271...279TADG-12 peptide 57 Ile Gln Val Gly Leu Val Ser Leu Leu 5 58 9 PRT Homosapiens 397...405 TADG-12 peptide 58 Cys Gln Gly Asp Ser Gly Gly Pro Leu5 59 9 PRT Homo sapiens 270...278 TADG-12 peptide 59 Thr Ile Gln Val GlyLeu Val Ser Leu 5 60 9 PRT Homo sapiens 56...64 TADG-12 peptide 60 GlyIle Ile Ala Leu Ile Leu Ala Leu 5 61 9 PRT Homo sapiens 110...118TADG-12 peptide 61 Arg Val Gly Gly Gln Asn Ala Val Leu 5 62 9 PRT Homosapiens 217...225 TADG-12 peptide 62 Arg Ile Val Gly Gly Asn Met Ser Leu5 63 9 PRT Homo sapiens 130...138 TADG-12 peptide 63 Cys Ser Asp Asp TrpLys Gly His Tyr 5 64 9 PRT Homo sapiens 8...16 TADG-12 peptide 64 AlaVal Glu Ala Pro Phe Ser Phe Arg 5 65 9 PRT Homo sapiens 328...336TADG-12 peptide 65 Asn Ser Glu Glu Asn Phe Pro Asp Gly 5 66 9 PRT Homosapiens 3...11 TADG-12 peptide 66 Glu Asn Asp Pro Pro Ala Val Glu Ala 567 9 PRT Homo sapiens 98...106 TADG-12 peptide 67 Asp Cys Lys Asp GlyGlu Asp Glu Tyr 5 68 9 PRT Homo sapiens 346...354 TADG-12 peptide 68 AlaThr Glu Asp Gly Gly Asp Ala Ser 5 69 9 PRT Homo sapiens 360...368TADG-12 peptide 69 Ala Ala Val Pro Leu Ile Ser Asn Lys 5 70 9 PRT Homosapiens 153...161 TADG-12 peptide 70 Ser Ser Asp Asn Leu Arg Val Ser Ser5 71 9 PRT Homo sapiens 182...190 TADG-12 peptide 71 Val Thr Ala Leu HisHis Ser Val Tyr 5 72 9 PRT Homo sapiens 143...151 TADG-12 peptide 72 CysAla Gln Leu Gly Phe Pro Ser Tyr 5 73 9 PRT Homo sapiens 259...267TADG-12 peptide 73 Cys Val Tyr Asp Leu Tyr Leu Pro Lys 5 74 9 PRT Homosapiens 369...377 TADG-12 peptide 74 Ile Cys Asn His Arg Asp Val Tyr Gly5 75 9 PRT Homo sapiens 278...286 TADG-12 peptide 75 Leu Leu Asp Asn ProAla Pro Ser His 5 76 9 PRT Homo sapiens 426...434 TADG-12 peptide 76 CysAla Glu Val Asn Lys Pro Gly Val 5 77 9 PRT Homo sapiens 32...40 TADG-12peptide 77 Asp Ala Asp Ala Val Ala Ala Gln Ile 5 78 9 PRT Homo sapiens406...414 TADG-12 peptide 78 Val Cys Gln Glu Arg Arg Leu Trp Lys 5 79 9PRT Homo sapiens 329...337 TADG-12 peptide 79 Ser Glu Glu Asn Phe ProAsp Gly Lys 5 80 9 PRT Homo sapiens 303...311 TADG-12 peptide 80 Gly AsnAsp Ile Ala Leu Met Lys Leu 5 81 9 PRT Homo sapiens 127...135 TADG-12peptide 81 Lys Thr Met Cys Ser Asp Asp Trp Lys 5 82 9 PRT Homo sapiens440...448 TADG-12 peptide 82 Phe Leu Asp Trp Ile His Glu Gln Met 5 83 9PRT Homo sapiens 433...441 TADG-12 peptide 83 Val Tyr Thr Arg Val ThrSer Phe Leu 5 84 9 PRT Homo sapiens 263...271 TADG-12 peptide 84 Leu TyrLeu Pro Lys Ser Trp Thr Ile 5 85 9 PRT Homo sapiens 169...177 TADG-12peptide 85 Glu Phe Val Ser Ile Asp His Leu Leu 5 86 9 PRT Homo sapiens296...304 TADG-12 peptide 86 Lys Tyr Lys Pro Lys Arg Leu Gly Asn 5 87 9PRT Homo sapiens 16...24 TADG-12 peptide 87 Arg Ser Leu Phe Gly Leu AspAsp Leu 5 88 9 PRT Homo sapiens 267...275 TADG-12 peptide 88 Lys Ser TrpThr Ile Gln Val Gly Leu 5 89 9 PRT Homo sapiens 81...89 TADG-12 peptide89 Arg Ser Ser Phe Lys Cys Ile Glu Leu 5 90 9 PRT Homo sapiens 375...383TADG-12 peptide 90 Val Tyr Gly Gly Ile Ile Ser Pro Ser 5 91 9 PRT Homosapiens 110...118 TADG-12 peptide 91 Arg Val Gly Gly Gln Asn Ala Val Leu5 92 9 PRT Homo sapiens 189...197 TADG-12 peptide 92 Val Tyr Val Arg GluGly Cys Ala Ser 5 93 9 PRT Homo sapiens 165...173 TADG-12 peptide 93 GlnPhe Arg Glu Glu Phe Val Ser Ile 5 94 9 PRT Homo sapiens 10...18 TADG-12peptide 94 Glu Ala Pro Phe Ser Phe Arg Ser Leu 5 95 9 PRT Homo sapiens407...415 TADG-12 peptide 95 Cys Gln Glu Arg Arg Leu Trp Lys Leu 5 96 9PRT Homo sapiens 381...389 TADG-12 peptide 96 Ser Pro Ser Met Leu CysAla Gly Tyr 5 97 9 PRT Homo sapiens 375...383 TADG-12 peptide 97 Val TyrGly Gly Ile Ile Ser Pro Ser 5 98 9 PRT Homo sapiens 381...389 TADG-12peptide 98 Ser Pro Ser Met Leu Cys Ala Gly Tyr 5 99 9 PRT Homo sapiens362...370 TADG-12 peptide 99 Val Pro Leu Ile Ser Asn Lys Ile Cys 5 100 9PRT Homo sapiens 373...381 TADG-12 peptide 100 Arg Asp Val Tyr Gly GlyIle Ile Ser 5 101 9 PRT Homo sapiens 283...291 TADG-12 peptide 101 AlaPro Ser His Leu Val Glu Lys Ile 5 102 9 PRT Homo sapiens 177...185TADG-12 peptide 102 Leu Pro Asp Asp Lys Val Thr Ala Leu 5 103 9 PRT Homosapiens 47...55 TADG-12 peptide 103 Glu Val Phe Ser Gln Ser Ser Ser Leu5 104 9 PRT Homo sapiens 36...44 TADG-12 peptide 104 Val Ala Ala Gln IleLeu Ser Leu Leu 5 105 9 PRT Homo sapiens 255...263 TADG-12 peptide 105Thr Ala Ala His Cys Val Tyr Asp Leu 5 106 9 PRT Homo sapiens 138...146TADG-12 peptide 106 Tyr Ala Asn Val Ala Cys Ala Gln Leu 5 107 9 PRT Homosapiens 195...203 TADG-12 peptide 107 Cys Ala Ser Gly His Val Val ThrLeu 5 108 9 PRT Homo sapiens 215...223 TADG-12 peptide 108 Ser Ser ArgIle Val Gly Gly Asn Met 5 109 9 PRT Homo sapiens 298...306 TADG-12peptide 109 Lys Pro Lys Arg Leu Gly Asn Asp Ile 5 110 9 PRT Homo sapiens313...321 TADG-12 peptide 110 Gly Pro Leu Thr Phe Asn Glu Met Ile 5 1119 PRT Homo sapiens 108...116 TADG-12 peptide 111 Cys Val Arg Val Gly GlyGln Asn Ala 5 112 9 PRT Homo sapiens 294...302 TADG-12 peptide 112 HisSer Lys Tyr Lys Pro Lys Arg Leu 5 113 9 PRT Homo sapiens 265...273TADG-12 peptide 113 Leu Pro Lys Ser Trp Thr Ile Gln Val 5 114 9 PRT Homosapiens 88...96 TADG-12 peptide 114 Glu Leu Ile Thr Arg Cys Asp Gly Val5 115 9 PRT Homo sapiens 79...87 TADG-12 peptide 115 Arg Cys Arg Ser SerPhe Lys Cys Ile 5 116 9 PRT Homo sapiens 255...263 TADG-12 peptide 116Thr Ala Ala His Cys Val Tyr Asp Leu 5 117 9 PRT Homo sapiens 207...215TADG-12 peptide 117 Ala Cys Gly His Arg Arg Gly Tyr Ser 5 118 9 PRT Homosapiens 154...162 TADG-12 peptide 118 Ser Asp Asn Leu Arg Val Ser SerLeu 5 119 9 PRT Homo sapiens 300...308 TADG-12 peptide 119 Lys Arg LeuGly Asn Asp Ile Ala Leu 5 120 9 PRT Homo sapiens 435...443 TADG-12peptide 120 Thr Arg Val Thr Ser Phe Leu Asp Trp 5 121 9 PRT Homo sapiens376...384 TADG-12 peptide 121 Tyr Gly Gly Ile Ile Ser Pro Ser Met 5 1229 PRT Homo sapiens 410...418 TADG-12 peptide 122 Arg Arg Leu Trp Lys LeuVal Gly Ala 5 123 9 PRT Homo sapiens 210...218 TADG-12 peptide 123 HisArg Arg Gly Tyr Ser Ser Arg Ile 5 124 9 PRT Homo sapiens 109...117TADG-12 peptide 124 Val Arg Val Gly Gly Gln Asn Ala Val 5 125 9 PRT Homosapiens 191...199 TADG-12 peptide 125 Val Arg Glu Gly Cys Ala Ser GlyHis 5 126 9 PRT Homo sapiens 78...86 TADG-12 peptide 126 Tyr Arg Cys ArgSer Ser Phe Lys Cys 5 127 9 PRT Homo sapiens 113...121 TADG-12 peptide127 Gly Gln Asn Ala Val Leu Gln Val Phe 5 128 9 PRT Homo sapiens 91...99TADG-12 peptide 128 Thr Arg Cys Asp Gly Val Ser Asp Cys 5 129 9 PRT Homosapiens 38...46 TADG-12 peptide 129 Ala Gln Ile Leu Ser Leu Leu Pro Phe5 130 9 PRT Homo sapiens 211...219 TADG-12 peptide 130 Arg Arg Gly TyrSer Ser Arg Ile Val 5 131 9 PRT Homo sapiens 216...224 TADG-12 peptide131 Ser Arg Ile Val Gly Gly Asn Met Ser 5 132 9 PRT Homo sapiens118...126 TADG-12 peptide 132 Leu Gln Val Phe Thr Ala Ala Ser Trp 5 1339 PRT Homo sapiens 370...378 TADG-12 peptide 133 Cys Asn His Arg Asp ValTyr Gly Gly 5 134 9 PRT Homo sapiens 393...401 TADG-12 peptide 134 GlyVal Asp Ser Cys Gln Gly Asp Ser 5 135 9 PRT Homo sapiens 235...243TADG-12 peptide 135 Leu Gln Phe Gln Gly Tyr His Leu Cys 5 136 9 PRT Homosapiens 427...435 TADG-12 peptide 136 Ala Glu Val Asn Lys Pro Gly ValTyr 5 137 9 PRT Homo sapiens 162...170 TADG-12 peptide 137 Leu Glu GlyGln Phe Arg Glu Glu Phe 5 138 9 PRT Homo sapiens 9...17 TADG-12 peptide138 Val Glu Ala Pro Phe Ser Phe Arg Ser 5 139 9 PRT Homo sapiens318...326 TADG-12 peptide 139 Asn Glu Met Ile Gln Pro Val Cys Leu 5 1409 PRT Homo sapiens 256...264 TADG-12 peptide 140 Ala Ala His Cys Val TyrAsp Leu Tyr 5 141 9 PRT Homo sapiens 46...54 TADG-12 peptide 141 Phe GluVal Phe Ser Gln Ser Ser Ser 5 142 9 PRT Homo sapiens 64...72 TADG-12peptide 142 Leu Ala Ile Gly Leu Gly Ile His Phe 5 143 9 PRT Homo sapiens192...200 TADG-12 peptide 143 Arg Glu Gly Cys Ala Ser Gly His Val 5 1449 PRT Homo sapiens 330...338 TADG-12 peptide 144 Glu Glu Asn Phe Pro AspGly Lys Val 5 145 9 PRT Homo sapiens 182...190 TADG-12 peptide 145 ValThr Ala Leu His His Ser Val Tyr 5 146 9 PRT Homo sapiens 408...416TADG-12 peptide 146 Gln Glu Arg Arg Leu Trp Lys Leu Val 5 147 9 PRT Homosapiens 206...214 TADG-12 peptide 147 Thr Ala Cys Gly His Arg Arg GlyTyr 5 148 9 PRT Homo sapiens 5...13 TADG-12 peptide 148 Asp Pro Pro AlaVal Glu Ala Pro Phe 5 149 9 PRT Homo sapiens 261...269 TADG-12 peptide149 Tyr Asp Leu Tyr Leu Pro Lys Ser Trp 5 150 9 PRT Homo sapiens 33...41TADG-12 peptide 150 Ala Asp Ala Val Ala Ala Gln Ile Leu 5 151 9 PRT Homosapiens 168...176 TADG-12 peptide 151 Glu Glu Phe Val Ser Ile Asp HisLeu 5 152 9 PRT Homo sapiens 304...312 TADG-12 peptide 152 Asn Asp IleAla Leu Met Lys Leu Ala 5 153 9 PRT Homo sapiens 104...112 TADG-12peptide 153 Asp Glu Tyr Arg Cys Val Arg Val Gly 5 154 344 PRT Homosapiens complete amino acid sequence of TADG-12D 154 Met Gly Glu Asn AspPro Pro Ala Val Glu Ala Pro Phe Ser Phe 5 10 15 Arg Ser Leu Phe Gly LeuAsp Asp Leu Lys Ile Ser Pro Val Ala 20 25 30 Pro Asp Ala Asp Ala Val AlaAla Gln Ile Leu Ser Leu Leu Pro 35 40 45 Leu Lys Phe Phe Pro Ile Ile ValIle Gly Ile Ile Ala Leu Ile 50 55 60 Leu Ala Leu Ala Ile Gly Leu Gly IleHis Phe Asp Cys Ser Gly 65 70 75 Lys Tyr Arg Cys Arg Ser Ser Phe Lys CysIle Glu Leu Ile Ala 80 85 90 Arg Cys Asp Gly Val Ser Asp Cys Lys Asp GlyGlu Asp Glu Tyr 95 100 105 Arg Cys Val Arg Val Gly Gly Gln Asn Ala ValLeu Gln Val Phe 110 115 120 Thr Ala Ala Ser Trp Lys Thr Met Cys Ser AspAsp Trp Lys Gly 125 130 135 His Tyr Ala Asn Val Ala Cys Ala Gln Leu GlyPhe Pro Ser Tyr 140 145 150 Val Ser Ser Asp Asn Leu Arg Val Ser Ser LeuGlu Gly Gln Phe 155 160 165 Arg Glu Glu Phe Val Ser Ile Asp His Leu LeuPro Asp Asp Lys 170 175 180 Val Thr Ala Leu His His Ser Val Tyr Val ArgGlu Gly Cys Ala 185 190 195 Ser Gly His Val Val Thr Leu Gln Cys Thr AlaCys Gly His Arg 200 205 210 Arg Gly Tyr Ser Ser Arg Ile Val Gly Gly AsnMet Ser Leu Leu 215 220 225 Ser Gln Trp Pro Trp Gln Ala Ser Leu Gln PheGln Gly Tyr His 230 235 240 Leu Cys Gly Gly Ser Val Ile Thr Pro Leu TrpIle Ile Thr Ala 245 250 255 Ala His Cys Val Tyr Asp Leu Tyr Leu Pro LysSer Trp Thr Ile 260 265 270 Gln Val Gly Leu Val Ser Leu Leu Asp Asn ProAla Pro Ser His 275 280 285 Leu Val Glu Lys Ile Val Tyr His Ser Lys TyrLys Pro Lys Arg 290 295 300 Leu Gly Asn Asp Ile Ala Leu Met Lys Leu AlaGly Pro Leu Thr 305 310 315 Phe Asn Gly Thr Ser Gly Ser Leu Cys Gly SerAla Ala Leu Pro 320 325 330 Leu Phe Gln Glu Asp Leu Gln Leu Leu Ile GluAla Phe Leu 335 340 344 155 1359 DNA Homo sapiens CDS entire cDNAsequence of TADG-12D 155 accgggcacc ggacggctcg ggtactttcg ttcttaattaggtcatgccc 50 gtgtgagcca ggaaagggct gtgtttatgg gaagccagta acactgtggc 100ctactatctc ttccgtggtg ccatctacat ttttgggact cgggaattat 150 gaggtagaggtggaggcgga gccggatgtc agaggtcctg aaatagtcac 200 catgggggaa aatgatccgcctgctgttga agcccccttc tcattccgat 250 cgctttttgg ccttgatgat ttgaaaataagtcctgttgc accagatgca 300 gatgctgttg ctgcacagat cctgtcactg ctgccattgaagttttttcc 350 aatcatcgtc attgggatca ttgcattgat attagcactg gccattggtc400 tgggcatcca cttcgactgc tcagggaagt acagatgtcg ctcatccttt 450aagtgtatcg agctgatagc tcgatgtgac ggagtctcgg attgcaaaga 500 cggggaggacgagtaccgct gtgtccgggt gggtggtcag aatgccgtgc 550 tccaggtgtt cacagctgcttcgtggaaga ccatgtgctc cgatgactgg 600 aagggtcact acgcaaatgt tgcctgtgcccaactgggtt tcccaagcta 650 tgtgagttca gataacctca gagtgagctc gctggaggggcagttccggg 700 aggagtttgt gtccatcgat cacctcttgc cagatgacaa ggtgactgca750 ttacaccact cagtatatgt gagggaggga tgtgcctctg gccacgtggt 800taccttgcag tgcacagcct gtggtcatag aaggggctac agctcacgca 850 tcgtgggtggaaacatgtcc ttgctctcgc agtggccctg gcaggccagc 900 cttcagttcc agggctaccacctgtgcggg ggctctgtca tcacgcccct 950 gtggatcatc actgctgcac actgtgtttatgacttgtac ctccccaagt 1000 catggaccat ccaggtgggt ctagtttccc tgttggacaatccagcccca 1050 tcccacttgg tggagaagat tgtctaccac agcaagtaca agccaaagag1100 gctgggcaat gacatcgccc ttatgaagct ggccgggcca ctcacgttca 1150atggtacatc tgggtctcta tgtggttctg cagctcttcc tttgtttcaa 1200 gaggatttgcaattgctcat tgaagcattc ttatgatggc tgctttataa 1250 tccttgtcag atattaataattccaactcc tgattcatgt tggtgttggc 1300 atcagttgat tatcttttct cattaaaattgtgatgctcc taaaaaaaaa 1350 aaaaaaaaa 1359 156 234 PRT Homo sapiensDOMAIN protease domain of TADG-12 156 Asn Met Ser Leu Leu Ser Gln TrpPro Trp Gln Ala Ser Leu Gln 5 10 15 Phe Gln Gly Tyr His Leu Cys Gly GlySer Val Ile Thr Pro Leu 20 25 30 Trp Ile Ile Thr Ala Ala His Cys Val TyrAsp Leu Tyr Leu Pro 35 40 45 Lys Ser Trp Thr Ile Gln Val Gly Leu Val SerLeu Leu Asp Asn 50 55 60 Pro Ala Pro Ser His Leu Val Glu Lys Ile Val TyrHis Ser Lys 65 70 75 Tyr Lys Pro Lys Arg Leu Gly Asn Asp Ile Ala Leu MetLys Leu 80 85 90 Ala Gly Pro Leu Thr Phe Asn Glu Met Ile Gln Pro Val CysLeu 95 100 105 Pro Asn Ser Glu Glu Asn Phe Pro Asp Gly Lys Val Cys TrpThr 110 115 120 Ser Gly Trp Gly Ala Thr Glu Asp Gly Ala Gly Asp Ala SerPro 125 130 135 Val Leu Asn His Ala Ala Val Pro Leu Ile Ser Asn Lys IleCys 140 145 150 Asn His Arg Asp Val Tyr Gly Gly Ile Ile Ser Pro Ser MetLeu 155 160 165 Cys Ala Gly Tyr Leu Thr Gly Gly Val Asp Ser Cys Gln GlyAsp 170 175 180 Ser Gly Gly Pro Leu Val Cys Gln Glu Arg Arg Leu Trp LysLeu 185 190 195 Val Gly Ala Thr Ser Phe Gly Ile Gly Cys Ala Glu Val AsnLys 200 205 210 Pro Gly Val Tyr Thr Arg Val Thr Ser Phe Leu Asp Trp IleHis 215 220 225 Glu Gln Met Glu Arg Asp Leu Lys Thr 230 234 157 124 PRTHomo sapiens DOMAIN protease domain of TADG-12D 157 Asn Met Ser Leu LeuSer Gln Trp Pro Trp Gln Ala Ser Leu Gln 5 10 15 Phe Gln Gly Tyr His LeuCys Gly Gly Ser Val Ile Thr Pro Leu 20 25 30 Trp Ile Ile Thr Ala Ala HisCys Val Tyr Asp Leu Tyr Leu Pro 35 40 45 Lys Ser Trp Thr Ile Gln Val GlyLeu Val Ser Leu Leu Asp Asn 50 55 60 Pro Ala Pro Ser His Leu Val Glu LysIle Val Tyr His Ser Lys 65 70 75 Tyr Lys Pro Lys Arg Leu Gly Asn Asp IleAla Leu Met Lys Leu 80 85 90 Ala Gly Pro Leu Thr Phe Asn Gly Thr Ser GlySer Leu Cys Gly 95 100 105 Ser Ala Ala Leu Pro Leu Phe Gln Glu Asp LeuGln Leu Leu Ile 110 115 120 Glu Ala Phe Leu 124 158 73 PRT Homo sapiensDOMAIN protease domain of TADG-12V 158 Asn Met Ser Leu Leu Ser Gln TrpPro Trp Gln Ala Ser Leu Gln 5 10 15 Phe Gln Gly Tyr His Leu Cys Gly GlySer Val Ile Thr Pro Leu 20 25 30 Trp Ile Ile Thr Ala Ala His Cys Val TyrGlu Ile Val Ala Pro 35 40 45 Arg Glu Arg Ala Asp Arg Arg Gly Arg Lys LeuLeu Cys Trp Arg 50 55 60 Lys Pro Thr Lys Met Lys Gly Pro Arg Pro Ser HisSer 65 70 73

What is claimed is:
 1. A DNA fragment encoding a Tumor AssociatedDifferentially-Expressed Gene-12 (TADG-12) protein or a splicing variantof TADG-12, said DNA is selected from the group consisting of: (a) anisolated DNA fragment encoding a TADG-12 protein that has the amino acidsequence of SEQ ID NO. 2; (b) an isolated DNA fragment encoding aTADG-12 splicing variant, said variant protein has an amino acidsequence comprising part of SEQ ID NO. 2; and (c) an isolated DNAfragment differing from the isolated DNA fragments of (a) and (b) abovein codon sequence due to the degeneracy of the genetic code, and whichencodes a TADG-12 protein or a splicing variant of TADG-12.
 2. The DNAfragment of claim 1, wherein said DNA fragment has the sequence selectedfrom the group consisting of SEQ ID NOs. 1, 3 and
 155. 3. The DNAfragment of claim 1, wherein said variant TADG-12 protein has an aminoacid sequence selected from the group consisting of SEQ ID NO. 4 and SEQID NO.
 154. 4. A vector comprising the DNA fragment of claim 1 andregulatory elements necessary for expressing said DNA in a cell.
 5. Thevector of claim 4, wherein said DNA fragment encodes a TADG-12 proteinhaving the amino acid sequence selected from the group consisting of SEQID NOs. 2, 4 and
 154. 6. A host cell transfected with the vector ofclaim 4, said vector expressing a TADG-12 protein.
 7. The host cell ofclaim 6, wherein said cell is selected from the group consisting of abacterial cell, a mammalian cell, a plant cell and an insect cell.
 8. Anantisense oligonucleotide complementary to the DNA fragment of claim 1.9. An isolated and purified TADG-12 protein encoded for by a DNAselected from the group consisting of: (a) isolated DNA encoding aTADG-12 protein that has the amino acid sequence of SEQ ID NO. 2; (b)isolated DNA encoding a TADG-12 splicing variant, said variant proteinhas an amino acid sequence comprising part of SEQ ID NO. 2; and (c)isolated DNA differing from the isolated DNA of (a) and (b) above incodon sequence due to the degeneracy of the genetic code, and whichencodes a TADG-12 protein or a splicing variant of TADG-12.
 10. Theisolated and purified TADG-12 protein of claim 9, wherein said TADG-12protein has an amino acid sequence selected from the group consisting ofSEQ ID NOs. 2, 4 and
 154. 11. An antibody directed against the TADG-12protein of claim
 9. 12. A method for detecting malignant hyperplasia ina biological sample, comprising the steps of: (a) isolating mRNA fromsaid sample; and (b) detecting in said sample mRNA encoding a TADG-12protein or a variant thereof, wherein the presence of said mRNA in saidsample is indicative of the presence of malignant hyperplasia, whereinthe absence of said mRNA in said sample is indicative of the absence ofmalignant hyperplasia.
 13. The method of claim 12, wherein saiddetection of TADG-12 mRNA is by a method selected from the groupconsisting of PCR amplification, Northern blot, dot blot and a DNA arraychip.
 14. The method of claim 13, wherein said PCR amplification usesprimers selected from the group consisting of SEQ ID Nos. 28-31.
 15. Themethod of claim 12, wherein said TADG-12 protein has an amino acidsequence selected from the group consisting of SEQ ID NOs. 2, 4 and 154.16. The method of claim 12, wherein said biological sample is selectedfrom the group consisting of blood, urine, saliva, tears, interstitialfluid, ascites fluid, tumor tissue biopsy and circulating tumor cells.17. A method for detecting malignant hyperplasia in a biological sample,comprising the steps of: (a) isolating protein from said sample; and (b)detecting in said sample a TADG-12 protein or a variant thereof, whereinthe presence of said TADG-12 protein in said sample is indicative of thepresence of malignant hyperplasia, wherein the absence of said TADG-12protein in said sample is indicative of the absence of malignanthyperplasia.
 18. The method of claim 17, wherein said detection is b y amethod selected from the group consisting of Western blot, dot blot,ELISA sandwich assay, radioimmunoassay and flow cytometry analysis. 19.The method of claim 17, wherein said TADG-12 protein has an amino acidsequence selected from the group consisting of SEQ ID NOs. 2, 4 and 154.20. The method of claim 17, wherein said biological sample is selectedfrom the group consisting of blood, urine, saliva, tears, interstitialfluid, ascites fluid, tumor tissue biopsy and circulating tumor cells.21. A method of inhibiting expression of endogenous TADG-12 mRNA in acell, comprising the step of: introducing a vector into a cell, whereinsaid vector comprises a DNA fragment of TADG-12 in opposite orientationoperably linked to elements necessary for expression, wherein expressionof said vector in said cell produces TADG-12 antisense mRNA, whereinsaid TADG-12 antisense mRNA hybridizes to endogenous TADG-12 mRNA,thereby inhibiting expression of endogenous TADG-12 mRNA in said cell.22. A method of inhibiting expression of a TADG-12 protein in a cell,comprising the step of: introducing an antibody into a cell, whereinsaid antibody is directed against a TADG-12 protein or fragment thereof,wherein binding of said antibody to said TADG-12 protein or fragmentthereof inhibits expression of said TADG-12 protein.
 23. A method oftargeted therapy to an individual, comprising the step of: administeringa compound to an individual, wherein said compound has a targetingmoiety and a therapeutic moiety, wherein said targeting moiety isspecific for a TADG-12 protein.
 24. The method of claim 23, wherein saidtargeting moiety is selected from the group consisting of an antibodydirected against a TADG-12 protein and a ligand or ligand binding domainthat binds a TADG-12 protein.
 25. The method of claim 23, wherein saidTADG-12 protein has an amino acid sequence selected from the groupconsisting of SEQ ID NOs. 2, 4 and
 154. 26. The method of claim 23,wherein said therapeutic moiety is selected from the group consisting ofa radioisotope, a toxin, a chemotherapeutic agent, an immune stimulantand a cytotoxic agent.
 27. The method of claim 23, wherein saidindividual suffers from a disease selected from the group consisting ofovarian cancer, lung cancer, prostate cancer, and colon cancer.
 28. Amethod of vaccinating an individual against TADG-12, comprising the stepof: inoculating the individual with a TADG-12 protein or fragmentthereof, wherein said inoculation elicits an immune response in saidindividual, thereby vaccinating said individual against TADG-12.
 29. Themethod of claim 28, wherein said individual has a cancer, is suspectedof having a cancer or is at risk of getting a cancer.
 30. The method ofclaim 28, wherein said TADG-12 protein has an amino acid sequenceselected from the group consisting of SEQ ID NOs. 2, 4 and
 154. 31. Themethod of claim 28, wherein said TADG-12 fragment has a sequence shownin SEQ ID NO.
 8. 32. The method of claim 28, wherein said TADG-12fragment is a 9-residue fragment selected from the group consisting ofSEQ ID NOs. 35, 36, 55, 56, 83, 84, 97, 98, 119, 120, 122, 123 and 136.33. An immunogenic composition, comprising a n immunogenic fragment of aTADG-12 protein and an appropriate adjuvant.
 34. The immunogeniccomposition of claim 33, wherein said immunogenic fragment of a TADG-12protein has a sequence shown in SEQ ID No.
 8. 35. The immunogeniccomposition of claim 33, wherein said immunogenic fragment of a TADG-12protein is a 9-residue fragment selected from the group consisting ofSEQ ID NOs. 35, 36, 55, 56, 83, 84, 97, 98, 119, 120, 122, 123 and 136.